Agilent 1290 Infinity LC
Injectors HTC/HTS
User Manual
Agilent Technologies
Notices
© Agilent Technologies, Inc. 2009,
2011-2012
No part of this manual may be reproduced
in any form or by any means (including electronic storage and retrieval or translation
into a foreign language) without prior agreement and written consent from Agilent
Technologies, Inc. as governed by United
States and international copyright laws.
Manual Part Number
G4277-90000 Rev. B
Edition
05/2012
Printed in Germany
Agilent Technologies
Hewlett-Packard-Strasse 8
76337 Waldbronn
This product may be used as a component of an in vitro diagnostic system if the system is registered with
the appropriate authorities and complies with the relevant regulations.
Otherwise, it is intended only for general laboratory use.
Warranty
The material contained in this document is provided “as is,” and is subject to being changed, without notice,
in future editions. Further, to the maximum extent permitted by applicable
law, Agilent disclaims all warranties,
either express or implied, with regard
to this manual and any information
contained herein, including but not
limited to the implied warranties of
merchantability and fitness for a particular purpose. Agilent shall not be
liable for errors or for incidental or
consequential damages in connection
with the furnishing, use, or performance of this document or of any
information contained herein. Should
Agilent and the user have a separate
written agreement with warranty
terms covering the material in this
document that conflict with these
terms, the warranty terms in the separate agreement shall control.
receive no greater than Restricted Rights as
defined in FAR 52.227-19(c)(1-2) (June
1987). U.S. Government users will receive
no greater than Limited Rights as defined in
FAR 52.227-14 (June 1987) or DFAR
252.227-7015 (b)(2) (November 1995), as
applicable in any technical data.
Safety Notices
CAUTION
A CAUTION notice denotes a
hazard. It calls attention to an
operating procedure, practice, or
the like that, if not correctly performed or adhered to, could
result in damage to the product
or loss of important data. Do not
proceed beyond a CAUTION
notice until the indicated conditions are fully understood and
met.
Technology Licenses
The hardware and/or software described in
this document are furnished under a license
and may be used or copied only in accordance with the terms of such license.
Restricted Rights Legend
If software is for use in the performance of a
U.S. Government prime contract or subcontract, Software is delivered and licensed as
“Commercial computer software” as
defined in DFAR 252.227-7014 (June 1995),
or as a “commercial item” as defined in FAR
2.101(a) or as “Restricted computer software” as defined in FAR 52.227-19 (June
1987) or any equivalent agency regulation
or contract clause. Use, duplication or disclosure of Software is subject to Agilent
Technologies’ standard commercial license
terms, and non-DOD Departments and
Agencies of the U.S. Government will
WA R N I N G
A WARNING notice denotes a
hazard. It calls attention to an
operating procedure, practice,
or the like that, if not correctly
performed or adhered to, could
result in personal injury or
death. Do not proceed beyond a
WARNING notice until the indicated conditions are fully understood and met.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
In This Book
In This Book
This manual covers the Agilent 1290 Infinity LC Injectors HTC/HTS
1 General Information
This chapter provides safety information and gives an introduction on how to
use this manual.
2 Operating Instructions
This chapter explains the operational parameters of the Agilent 1290 Infinity
LC Injectors HTC/HTS System.
3 Description and Installation
This chapter gives an overview and information about the installation of your
Agilent 1290 Infinity LC Injectors HTC/HTS.
4 Appendices
This chapter provides addition information.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
3
Contents
Contents
1 General Information
7
Safety Information 8
How to Use this Manual
2 Operating Instructions
14
15
Using the Control Terminal
Methods 20
Job and Job Queue 23
Utility Functions 27
Logfile 37
Info Functions 38
Setup Functions 42
3 Description and Installation
17
45
General System Overview 47
Installation 54
Object Positions 88
Injection Valve 105
Interfacing the LC Injector HTC/HTS to other Devices
PAL Software Program 118
Establishing Communication with Computer 120
Special Functions 126
Troubleshooting 146
Replacing Parts 170
Maintaining the LC Injector HTC/HTS System 189
4
115
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Contents
4 Appendices
191
Definition of Terms 192
Conventions of Naming 195
Injection Cycles 196
DLW Principles and Operating Details
Software Flow Chart 246
External Connectors 253
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
200
5
Contents
6
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
1
General Information
Safety Information
8
How to Use this Manual
14
This chapter provides safety information and gives an introduction on how to
use this manual.
Agilent Technologies
7
1
General Information
Safety Information
Safety Information
General
Considerations
The LC Injector HTC/HTS System User Manual and the corresponding
“Addendum” for a specific module must be consulted by the user under all
circumstances before a unit is put in use.
Changes or modifications to this unit not expressly approved by the party
responsible for compliance could void the user’s authority to operate the
equipment.
The user shall be made aware that if the equipment is used in a manner not
specified by the manufacturer, the protection provided by the equipment may
be impaired.
When using the LC Injector HTC/HTS System, follow the generally accepted
procedures for quality control and methods development.
When you use the LC Injector HTC/HTS System in the field of
chromatographic analysis and you observe a change in the retention of a
particular compound, in the resolution between two compounds, or in peak
shape, immediately determine the reason for the changes. Until you determine
the cause of a change, do not rely on the separation results.
Electrical
Hazards
Every analytical instrument has specific hazards, so be sure to read and
comply with the following precautions. They will help ensure the safe,
long-term use of your LC Injector HTC/HTS System.
The Installation Category (Over voltage Category) for this instrument is Level
II. The Level II Category pertains to equipment that receives its electrical
power from the local level, such as an electrical wall outlet.
Only use fuses of the type and current rating specified. Do not use repaired
fuses and do not short-circuit the fuse holder.
The supplied power cord must be inserted into a power outlet with a
protective earth contact (ground). When using an extension cord, make sure
that the cord also has an earth contact.
8
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
General Information
Safety Information
1
Do not change the external or internal grounding connections. Tampering
with or disconnecting these connections could endanger you and/or damage
the LC Injector HTC/HTS System.
The instrument is properly grounded in accordance with these regulations
when shipped. You do not need to make any changes to the electrical
connections or the instrument's chassis to ensure safe operation.
The combination of a LC Injector HTC/HTS System with a LC/MS System does
require the safety measure as described by the LC/MS System manufacturer.
Detailed instructions for the safety grounding on the LC/MS system are
outlined in the corresponding operating/installation manual.
Agilent Technologies recommends to use a grounding cable connected on one
side at the Injection Valve, Loop or any other suitable direct metallic contact
and the other side at an appropriate grounding point at the LC/MS System.
This supplementary grounding measure will support the safety strategy of the
LC/MS System manufacturer.
Do not turn the instrument on if you suspect that it has incurred any kind of
electrical damage. Instead disconnect the power cord and contact a Agilent
Technologies representative for a product evaluation. Do not attempt to use
the instrument until it has been evaluated. Electrical damage may have
occurred if the LC Injector HTC/HTS System shows visible signs of damage,
exposure to any liquids or has been transported under severe stress.
Damage can also result if the instrument is stored for prolonged periods under
unfavorable conditions (e.g. subjected to heat, water, etc.).
In any case disconnect the power cord(s) from the power supply or from the
different power supplies if optional devices are installed before attempting
any type of maintenance.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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1
General Information
Safety Information
Capacitors inside the instrument may still be charged even if the instrument is
turned off.
To avoid damaging electrical parts, do not disconnect an electrical assembly
while power is applied to the LC Injector HTC/HTS system. Once the power is
turned off, wait approximately 30 seconds before you disconnect an assembly.
The instrument includes a number of integrated circuits. These circuits may
be damaged if exposed to excessive line voltage fluctuations and/or power
surges.
Never try to repair or replace any components of the instrument that is not
described in this manual without the assistance of a Agilent Technologies
representative.
There are no operator-serviceable or replaceable parts inside the power
supply(ies) or in the LC Injector HTC/HTS System. If a power supply is not
functioning, contact a Agilent Technologies representative.
The power supply for the LC Injector HTC/HTS Instrument has the symbols
1/0 on the label for the power switch to switch ON/OFF.
Any additional power supply for other devices like, Cooled Stack or a Valve
Module shows the symbols as shown below on the label for the power switch:
The symbols shall warn the user that in a emergency case more than one
power supply has to be turned OFF or more than one power cord has to be
pulled from power supply or from the wall outlet to shut down the complete
LC Injector HTC/HTS System.
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Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
General Information
Safety Information
1
If the basic LC Injector HTC/HTS System is installed, than a single power
supply is installed only. Turning OFF the power supply or pulling this single
power cord in an emergency case will stop the complete LC Injector HTC/HTS
System.
It is important that the power supply (ies) are in a location where the power
ON and OFF switch is accessible and easy to operate, and where it is possible
to unplug the AC power cord from the power supply/wall outlet in case of
emergency.
Other Hazards
To avoid injury during LC Injector HTC/HTS System operation, keep your
hands away from the syringe.
Do not operate the LC Injector HTC/HTS System without the safety shield. The
safety shield must be installed for safe operation.
To avoid injury, observe safe laboratory practice when you handle solvents,
change tubing, or operate the LC Injector HTC/HTS System. Know the physical
and chemical properties of the solvents you use. See the Material Safety Sheets
from the manufacturer for the solvents in use.
Use caution when working with any polymer tubing under pressure:
• Always wear eye protection when near pressurized polymer tubing.
• Do not use polymer tubing that has been severely stressed or kinked.
• Do not use polymer tubing, in particular not PEEK or Tefzel tubing, with
Tetrahydrofuran (THF), Dimethylsulfoxid (DMSO), chlorinated organic
solvents, concentrated mineral acids, such as Nitric, Phosphoric or Sulfuric
acids, or any related compounds to above listings.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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1
General Information
Safety Information
Do not use vials without a sealing cap, microtiter or deepwell plates without a
plate seal. Vapor phase from organic solvents can be hazardous and
flammable. Acidic vapor phase can cause corrosion to critical mechanical
parts.
Disposal
Do not dispose of this equipment or parts thereof unsorted in municipal
waste. Follow local municipal waste ordinances for proper disposal provisions
to reduce the environmental impact of waste electrical and electronic
equipment (WEEE). European Union customers: Call your local customer
service representative responsible for the PAL System for complimentary
equipment pick-up and recycling.
Lithium battery
An onboard lithium battery buffers the electronic memories, when the
instrument is turned off. Replace it only with the same or equivalent type
recommended by the equipment manufacturer.
Battery: Panasonic VL 2330, soldered directly on the electronic board.
Discharged lithium batteries shall be disposed off locally according to national
waste disposal regulations for batteries.
There are no operator-serviceable parts on the electronic boards. If an
electronic board fails, contact a Agilent Technologies representative.
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Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
General Information
Safety Information
Table 1
Symbol
1
Commonly Used Symbols
Description
Caution or refer to User Manual
Caution, Risk of Needle-Stick Puncture
Caution, Hot Surface or High Temperature
Direct Current
Alternating Current
Protective Conductor Terminal, Ground
Fuse
I
Electrical Power ON.
Used with Main LC Injector HTC/HTS Power Supply.
O
Electrical Power OFF.
Used with Main LC Injector HTC/HTS Power Supply.
Electrical Power ON for Only Part of the System.
Used with Optional Device(s)
Electrical Power OFF for Only Part of the System.
Used with Optional Device(s)
Caution, Risk of Electrical shock (high voltage)
Disposal, Do not dispose in municipal waste. Follow local waste regulations to reduce electrical and
electronic waste (WEEE).
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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1
General Information
How to Use this Manual
How to Use this Manual
The manual is divided into three major sections
• “Operating Instructions” on page 15)
• “Description and Installation” on page 45)
• “Appendices” on page 191
The "LC Injector HTC/HTS Operating instructions" are intended for infrequent
LC Injector HTC/HTS users or new users that are experienced in using
automated systems to perform existing analytical methods.
NOTE
The LC Injector HTC/HTS must be installed and set up properly before the Operating
Instructions in Section A can be used.
Users who are installing a LC Injector HTC/HTS system, LC Injector HTC/HTS
accessories or who need to make adjustments to an installed system should
consult "LC Injector HTC/HTS Description and Installation" in Section B.
The Appendices provide useful information such as the Software Flow Chart,
Definition of Terms or the LC Injector HTC/HTS Accessories guide.
14
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
2
Operating Instructions
Using the Control Terminal 17
Menu Screens 18
Function Keys 19
ESCape and STOP Keys 19
Scroll Knob and ENTER Button
19
Methods 20
Creating Methods 20
Edit / View Methods 22
Delete Methods 22
Job and Job Queue 23
Building and Starting a Job Queue 24
Aborting a Job Queue 26
Restarting an Aborted Job Queue 26
Utility Functions 27
Syringe 28
Tray 30
Injector 32
Wash Station 33
Vial 34
Dilutors 35
Tools 36
Logfile
37
Info Functions 38
Hardware 39
Software 39
Maintenance 40
Free Objects / Free Items
41
Agilent Technologies
15
2
Operating Instructions
How to Use this Manual
Setup Functions
Sounds 43
Time 43
Objects 44
42
This chapter explains the operational parameters of the Agilent 1290 Infinity LC
Injectors HTC/HTS System.
16
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Operating Instructions
Using the Control Terminal
2
Using the Control Terminal
The following procedures present the key steps required to set up and process
multiple groups of samples with the system. They are intended to provide an
overview for new users and a reminder for infrequent users. The system and
all accessories should be installed with Objects defined correctly. A syringe of
the specific type called for by a particular method should also be installed.
Figure 1 on page 17 illustrates the system control terminal and the
conventions used to enter, edit, and view information.
BZcji^iaZ
?dW
8jghdgWVg
]^\]a^\]ihVc^iZb^ca^hi
;jcXi^dc`ZnYZ[^c^i^dch
;jcXi^dc`Znh
EgZhh:H8VeZ
idgZijgcid
i]ZegZk^djhaZkZa
GdiViZi]ZdjiZg`cdWidbdkZi]ZXjghdgWVg
EgZhhi]Z^ccZg`cdWidhZaZXiVc^iZb]^\]a^\]iZYWni]ZXjghZgWVg#
EgZhhV\V^cid:CI:Gi]ZhZaZXi^dc#
EgZhhHIDEidVWdgideZgVi^dc
Figure 1
System Control Terminal and Conventions
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
17
2
Operating Instructions
Using the Control Terminal
Menu Screens
Different menu screens are displayed, depending on the system operating
status and the particular function being accessed by the operator. All menu
screens have the same basic format. The menu title is displayed at the top of
the screen. A list of items is displayed below the title. The date and time, or
status, are shown in the highlighted area above the Function key labels on the
bottom of the screen.
BZcj
BZi]dYh
A8&%%
HXgdaaYdlc
Figure 2
18
Accessing a Method Screen
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
2
Operating Instructions
Using the Control Terminal
Function Keys
Options for a particular menu are assigned to the corresponding function keys
(F1, F2, F3, F4) directly below each function key label.
Pressing the function key labeled as Home will always return to the Job Queue
menu.
ESCape and STOP Keys
Press ESCape to return to the previous menu. Press STOP to abort the current
Cycle, Job, or Job Queue.
Scroll Knob and ENTER Button
Rotate the outer knob to scroll through items in a menu list. To select a
highlighted item press the central knob (ENTER). Then use the outer knob to
scroll through available options for that item or to change a numeric value.
Then press the inner knob again to ENTER the displayed option. The inner
knob is also used for other operations that require an ENTER operation to
continue or complete an operation.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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2
Operating Instructions
Methods
Methods
Creating Methods
Methods can be defined by the user and assigned names up to eight characters
in length. Methods can be created, copied, edited, and viewed from the
Methods menu. Methods can be viewed (but not edited) from the Job Queue
menus.
Methods are created by either copying an existing Method or creating a new
Method.
To copy a Method, complete the sequence as follows:
1
BZcj
BZi]dYh
BZi]dYcVbZ
8denBZi]dY
You will be prompted to enter a name for the new Method.
2 Use the scroll knob and the left-right arrow function keys (F2 and F3) to
select among alphanumeric characters and spaces. Press the ENTER function
key (F4) to accept the name.
20
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Operating Instructions
Methods
2
To create a new Method, complete the following steps:
1
BZcj
BZi]dYh
HZaZXii]Z^chZgi^dced^ci
>chgiBZi]dY
2 Assign and enter a new Method name as above.
After a copy of the Method has been created, the Method parameters will
display and can be edited. The Cycle and Syringe entries cannot be changed.
3 If the Method is new (i.e. added), select and enter a Cycle that is
appropriate for the application.
4 Select the specific Syringe to be used by the Method.
NOTE
Once a Method has been created and saved, the Cycle and Syringe cannot be changed. To
use a different Cycle or Syringe, a new Method must be created.
5 Assign Parameter values according to the application requirements.
Consult ““LC-Inj” Cycle” on page 196 for details on specific items.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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2
Operating Instructions
Methods
Edit / View Methods
Method parameters (excluding Cycle and Syringe) can be viewed and
changed from the Method menu as follows:
Complete the following menu selections:
1
BZcj
BZi]dYh
BZi]dYcVbZ
:ciZg
2 Scroll to and select the parameter to be changed. Assign the new value and
press ENTER.
3 Exit from Parameter List by pressing either the Home function key (F4) to
return to the top-level Job Queue menu or ESCape to return to the previous
menu.
4 Method contents may be viewed from the Job Queue displays by selecting the
desired Job, pressing ENTER, followed by the View Method function key.
Delete Methods
Methods can be deleted from the Methods menu. Methods in use by an active
Job cannot be deleted.
1 Complete the following menu selections to delete a Method.
BZcj
BZi]dYh
BZi]dYcVbZ
9ZaBZi]dY
22
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Operating Instructions
Job and Job Queue
2
Job and Job Queue
A Job bundles the specified Tray with the designated vials (samples) and with
the Method to run those samples. Another term often used for “Job” in the
chromatographic field is “sequence”. If more than one Job is prepared, the
term Job Queue is used.
Before a Job can be activated the operator must verify that the Tray Type
matches the specified Tray and vial size (type). This step is done in Utilities
class.
BZcj
Ji^a^i^Zh
IgVn
hZaZXiIgVn
HXgdaaYdlc
Figure 3
Verifying Tray with corresponding Tray Type
1 Select the corresponding Tray Type. By activating F3 the injection unit can
be moved to the first position and to another two corner positions to verify
the correct selection of the Tray Type and teaching position.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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2
Operating Instructions
Job and Job Queue
Building and Starting a Job Queue
1 Power up the module. The JOB QUEUE screen is displayed.
Figure 4
Example Job Queue Screen
2 Load a sample Tray onto an available location in a Tray Holder or Stack.
Note the corresponding Tray name.
3 Add a new Job for the Tray. Press the Add Job key to bring up the default
Job.
4 For TRAY, select the Tray name (e.g. Stk1-01) that corresponds to the
location of the Tray that was just loaded.
5 Enter the First and Last sample number for this Job.
6 Select and enter the sample processing Method for this Job.
7 Press the Home function key (F4) to return to the JOB QUEUE screen.
24
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
2
Operating Instructions
Job and Job Queue
8 To add additional samples to be processed, repeat steps 2 - 7.
9 If necessary, replace and/or clean the syringe (liquid versions only). Press
the Menu key to see the available options for changing (F1 - Change Syringe)
and cleaning (F2 - Clean Syringe). To completely remove air bubbles, the
syringe should be primed manually.
10 If only one Job is to be processed, select the Job with the scroll knob. Press
Start key. In the dialog box "Select Job(s) to Process" select one of the
following options:
• All (Entire Job Queue starting from the top)
• Selected (Job selected with the cursor bar)
• Resume (Continue with the next Job after the one aborted)
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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2
Operating Instructions
Job and Job Queue
Aborting a Job Queue
1 Press STOP
2 Select one of the available options (Continue, Sample, Job, or Job Queue).
• Select Continue to resume processing with the current sample.
• Select Sample if there is a problem with only the current sample.
Processing will resume with the next sample.
• Select Job to abort processing all samples in the current Job. Processing
will resume with the next Job. The aborted Job is marked with an X.
• Select Job Queue to abort processing all Jobs. The JOB QUEUE screen will
be displayed. The aborted Job is marked with an X.
Restarting an Aborted Job Queue
1 Press the START key.
2 Select the Resume option. The Job after the last one marked as aborted will
be started.
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Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Operating Instructions
Utility Functions
2
Utility Functions
Utility functions, selectable from the Menu screen, provide quick access to
checking operations and parameters that may need to be changed. These
functions are available for the actual Syringe, Trays, Injectors, and the Wash
Station. They allow access to key functions without having to set up and
execute a Method and Job.
NOTE
If an item is used in the sample processing cycle, the appropriate Utility value will be
overwritten by the Method value.
BZcj
Ji^a^i^Zh
HXgdaaYdlc
Figure 5
Selecting Utilities Functions
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2
Operating Instructions
Utility Functions
Syringe
The following functions are available by pressing a Function Key:
Table 2
28
Syringe - Function Key
Function Key
Description
F1Chang Syr
The syringe is moved to a position in which the syringe assembly can be
completely lowered to facilitate removal of the syringe adapter. The syringe
can then be removed from the adapter and replaced. A prompt will be
displayed to specify the new syringe. The syringe must be installed before
pressing Enter.
F2Clean Syr
This Function is used to clean or prime the syringe prior to use. After
selecting F2 either Wash1 or Wash2 can be selected.
F3Set Pos
Set Pos is used to define the Chang Syr position
F4HOME
The Injection Unit moves to its HOME position and the Job Queue Menu is
displayed.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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Operating Instructions
Utility Functions
The following Syringe items may be changed by selecting the particular item:
Table 3
Syringe Items
Item
Description
Actual ID
Indicates the identification number (ID) of the currently inserted syringe. If
the syringe detection system is set to manual, the message Syringe: No
syringe is displayed.
Fill Volume
This parameter serves to control the filling of the syringe. It can occur that
air bubbles remain below the plunger after the first pull up. If the plunger is
moved up and down several times (see Fill Strokes), these air bubbles are
worked out. With this operation the syringe can be completely filled even
when using very small sample volumes.
Fill Strokes
Number of fill strokes. All fill strokes, except the last one, use the selected
fill volume. If the selected sample volume is higher than the fill volume, the
sample volume is used for all fill strokes. If zero is selected the plunger is
pulled up only once using the sample volume value.
Pullup Del
By using this item, a delay time can be selected between sample pullup and
ejection while filling the syringe. When the plunger reaches the zero
position during the fill strokes, the system waits half the Pullup Del time.
This allows for an air bubble to float away from the needle tip. This feature
is especially useful for removing any air bubble in the syringe and handling
viscous fluids.
Fill Speed
Speed of plunger movement used in all syringe filling operations.
Eject Speed
Speed of plunger movement used in all syringe eject operations except
sample injection.
Inject Speed
Speed of plunger movement for sample injection. Typically used for Fill
Strokes.
Plunger Chnge Pos
Plunger position during Change Syringe operation. The syringe plunger is
moved to a position where the syringe can be removed and replaced. The
value may be changed for different types of syringes.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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2
Operating Instructions
Utility Functions
Tray
After selecting the particular Tray to be accessed, the following functions are
available:
Table 4
30
Tray - Function Key
Function Key
Description
F2Block NdlG
F2 activates Needle Guide blocking. The option after activation is “Rel
NdlG”, releasing Needle Guide Blocking. It may be used to test the
functionality of the solenoid that blocks the needle guide.
F3Movto nnn
This function serves as a quick check to determine if the X,Y,Z-coordinates
are defined correctly for the selected Tray. To use this utility the selected
Tray, including the sample vials, must be present. After pressing “Movto
001” the Injection Unit moves to sample position no.1. This procedure can
be repeated for the last sample position in the first row and the last sample
position.
F4HOME
The Injection Unit moves to its HOME position and the Job Queue Menu is
displayed.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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Operating Instructions
Utility Functions
The following Tray items may be changed by selecting the particular item:
Table 5
Item
Description
Needle Penetr
Needle penetration depth into the sample vial. The needle penetration
depth for the selected Tray can be changed by entering the desired value.
Tray Type
The Tray Type which is selected for the Tray is shown. If the Tray enables
the use of different Tray Types it can be changed at this position.
Tray Offset X
If necessary, a correction to the ideal X-position of “Position 1” can be
made by using “Tray Offset X”.
Tray Offset Y
If necessary, a correction to the ideal Y-position of “Position 1” can be
made by using “Tray Offset Y”.
Tray Offset Z
If necessary, a correction to the ideal Z-position of “Position 1” can be
made by using “Tray Offset Z”.
dxRow1
Correction of any inclination of a Tray (plate) in the X-axis of a row (see
Figure 37 on page 90 for details).
dyRow1
Correction of any inclination of a Tray (plate) in the Y-axis of a row (see
Figure 37 on page 90 for details).
dzRow1
Correction of any inclination of a Tray (plate) in the Z-axis of a row (see
Figure 37 on page 90 for details).
dxCol1
Correction of any inclination of a Tray (plate) in the X-axis of a column (see
Figure 37 on page 90 for details).
dyCol1
Correction of any inclination of a Tray (plate) in the Y-axis of a column (see
Figure 37 on page 90 for details).
dzCol1
Correction of any inclination of a Tray (plate) in the Z-axis of a column (see
Figure 37 on page 90 for details).
1
NOTE
Tray - Items
Available beginning with PAL Firmware Level 4.1.X.
A “staggered pattern” is selectable in firmware class “Tray Type”. For details see “Tray
Type” on page 138.
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2
Operating Instructions
Utility Functions
Injector
After selecting the particular Injector to be accessed, the following functions
are available:
Table 6
Injector - Function Key
Function Key
Description
F3Movto Inj
The Injection Unit moves to the selected injector position. With this
function, e.g. the injectors LC Vlv1, Waste and Waste2 can be accessed.
By selecting the parameter “Needle Penetr” on the same screen, the
Injector Needle Penetration value can be checked or changed.
F4HOME
The Injection Unit moves to its HOME position and the Job Queue Menu is
displayed.
The following Injector item can be changed by selecting the particular item:
Table 7
32
Injector - Parameters
Item
Description
Needle Penetr
By selecting the parameter “Needle Penetr” the Injector Needle
Penetration value can be checked and/or changed. To ensure reproducible
sample injections and minimize carry-over it is critical that the needle
penetration depth be accurately set.
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Operating Instructions
Utility Functions
Wash Station
After selecting the specific Wash Station, the following Functions are
available:
Table 8
Wash Station - Function Key
Function Key
Description
F3Movto Wash
The injection unit moves to the selected Wash Station port. By selecting
the Parameter “Needle Penetr” on the same screen the Wash Station
Needle Penetration value can be checked or changed.
F4HOME
The Injection Unit moves to its HOME position and the Job Queue Menu is
displayed.
The following Wash Station Parameter may be changed by selecting the
particular item:
Table 9
Wash Station - Items
Item
Description
Needle Penetr
By selecting the Parameter “Needle Penetr” the Wash Station Needle
Penetration value can be checked and/or changed.
Rinse Time
If Rinse Time is activated (value > 0), the solenoid of a Fast or Active Wash
Station opens for the specified time after the syringe needle has been
removed from the wash port (after completion of the syringe wash cycle).
The solvent flows into the wash port without the restriction of the needle;
be aware of higher solvent consumption.
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Operating Instructions
Utility Functions
Vial
After selecting the specific Vial type, the following functions are available:
Table 10
Function Key
Description
F3Movto Vial
The injection unit moves to the selected vial type. By selecting the
parameter “Needle Penetr” on the same screen, the Wash Station Needle
Penetration value can be checked or changed.
F4HOME
The Injection Unit moves to its HOME position and the Job Queue Menu is
displayed.
Table 11
34
Vial - Function Key
Vial - Items
Item
Description
Needle Penetr
By selecting the parameter “Needle Penetr” the Wash Station needle
penetration value can be checked and/or changed.
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Operating Instructions
Utility Functions
Dilutors
After selecting the specific Dilutor, the following functions are available:
Table 12
Dilutors - Function Key
Function Key
Description
F1Prime
The Dilutor syringe is primed with solvent after activating the F1 function.
F2Chang DSyr
Activating F2 moves the plunger of the Dilutor Syringe to standby position
to allow easy access to change the syringe.
F4HOME
The Injection Unit moves to its HOME position and the Job Queue Menu is
displayed.
The following Dilutor items can be changed by selecting the particular item:
Table 13
Dilutor - Items
Item
Description
Syringe
Indicates the Dilutor side-port syringe inserted in the Z-axis. This function
allows selecting another syringe size.
Syr Dilut Pos
Activating this function moves the plunger of the side port syringe up by
the specified distance. This allows adjustment of the plunger tip of the side
port syringe exactly above the lower side port (solvent inlet). This fine
tuning is necessary after changing a side port syringe to allow unrestricted
solvent flow.
Dilutor Syr
Indicates Dilutor Syringe is installed. If the syringe size is changed, this
item must be adapted accordingly. The syringe-specific dimensions are
coordinated by this function.
Prime Volume
A volume to prime the Dilutor Syringe can be specified. The allowed range
can be fourfold higher than the actual syringe volume. This allows filling
and emptying a dilutor syringe more often than just once.
Pullup Delay
Using this item permits selecting a delay time between solvent filling and
ejection while filling (or priming) the syringe.
Fill Speed
Speed of plunger movement of Dilutor Syringe used in all syringe filling
operations.
Eject Speed
Speed of Dilutor Syringe plunger movement used in all dilutor syringe eject
operations.
Eject Delay
Using this item permits selecting a delay time between solvent ejection
and filling while filling (or priming) the syringe.
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Operating Instructions
Utility Functions
Tools
NOTE
The Object class Tools is available with Firmware version 3.0.X , 4.1.X. or higher.
After selecting the specific tool, the following functions become available:
Table 14
Tools - Function Key
Function Key
Description
F1Chk Offs
The item “Check Offset” can be used to verify the offset from the syringe
needle tip to the tool tip. This item is mainly used for the MALDI tool; it is
not active for the MHE tool.
F4HOME
The injection unit moves to its HOME position and the Job Queue Menu is
displayed.
The following tool items may be changed by selecting the particular item:
Table 15
36
Tools - Items
Item
Description
Teach Point
The reference point at which to teach the Object (Tool; MHETool) is
selectable. In the case of the MHETool, no extra position is necessary. The
Parking Station is the fix point. Select “None.”
ToolOffset X
If necessary, a correction to the ideal X-position of the MHETool can be
made by using “ToolOffsetXX”.
ToolOffset Y
If necessary, a correction to the ideal Y-position of the MHETool can be
made by using “ToolOffsetY”
ToolOffset Z
If necessary, a correction to the ideal Z-position of the MHETool can be
made by using “ToolOffsetZ” A “ToolOffsetZ” of -48 mm is necessary to
position the MHETool on the sample vial.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Operating Instructions
Logfile
2
Logfile
The logfile cannot be read-out directly on the terminal display.
However, F3 does allow a print out. A serial printer has to be connected to port
SER2. If a serial printer is not available use a serial/parallel converter.
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2
Operating Instructions
Info Functions
Info Functions
Info functions, selectable from the Menu screen, provide quick access to
information to be viewed. These info functions are available for the Hardware,
Software, Maintenance, Free Objects, and Free Items. All items with the exception
of “Maintenance” are read only.
BZcj
>c[d
HXgdaaYdlc
Figure 6
38
Selecting Info Functions
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Operating Instructions
Info Functions
2
Hardware
Table 16
Hardware
Item
Description
CPU SNo
The serial number (SNo.) of the PCB “APR CPU” is displayed.
CPU ID
Version number of the PCB “APR CPU”.
MOTIO ID
Version number of the PCB “APR CPU”.
Software
Table 17
Software
Item
Description
Firmware
Firmware version of System
Head Firmware
Firmware version of the Injection Unit
Terminal FW
Firmware version of the Terminal
Altera Firmware
Firmware version of the Altera component
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Operating Instructions
Info Functions
Maintenance
Table 18
NOTE
40
Maintenance
Item
Description
PlgStrokeCnt
This is a counter for syringe plunger movements. The actual number of
strokes is displayed. If the counter reaches the set limit (PlgStrkeLim) a
warning is displayed at the next start of a job (run). The system continues
but signals the user to verify syringe conditions. The counter can be set
back to zero to restart. There is only one counter for a syringe. If syringe
types are changed the system continues to count as if it were the same
type.
PlgStrokeLim
An upper limit for the syringe plunger strokes can be set.
Inject Count
This counter monitors the number of injections. The number of injection
valve switches is a helpful tool for the user to decide on a replacement of
parts of the injector system. The actual number of injections (valve
switches) is displayed. If the counter reaches the set limit (Inject Limit), a
warning is displayed at the next start of a job (run) (Inject Limit). The same
counter is used for injector penetrations with GC technique.
Inject Limit
An upper limit for the number of injections can be set.
Counters for the plunger movement and injector penetrations are available with LC Injector
HTC/HTS Firmware version 2.5.X or higher.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Operating Instructions
Info Functions
2
Free Objects / Free Items
In addition to the core software, the Firmware contains data for the Firmware
Objects. There are different classes of Objects, such as Syringes, Trays, Tray
Holders, etc. Each class of Objects contains Items. The items contain the actual
data such as X-, Y-, Z-positions.
The data are stored in a flash memory backed up by a battery. To optimize
RAM and Flash memory use, a certain section of memory has been reserved by
the software for each of the Objects and Object Items.
The percentage shown in the Info section provides an indication as to how
much of the reserved software space is still available.
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Operating Instructions
Setup Functions
Setup Functions
The Setup functions, selectable from the Menu screen, allow access to various
functions for the sampler system. The Sound, Time and Objects are basic
functions used at installation or if changes have been made over time.
BZcj
HZije
Figure 7
42
Selecting Setup Functions
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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Operating Instructions
Setup Functions
Sounds
Table 19
Sounds
Item
Description
Message Box
A specific dual beep tone signals that a pop-up window (Message Box) for
user intervention appears on the screen.
This beep signal can be turned on or off.
Warn Move
A beep sound is heard at the start of the module movement. It is advisable
to keep this function turned on for safety reasons.
End Cycle
A beep sound is heard at the end of a cycle. Select as desired.
End Job
A beep sound is heard at the end of a cycle. Select as desired.
Time
Table 20
Time - Item
Item
Description
Year
The year can be set for the module internal clock
Month
The month can be set for the module internal clock.
Day of Month
The day can be set for the module internal clock.
Hours
The hours can be set for the module internal clock.
Minutes
The minutes can be set for the module internal clock.
Seconds
The seconds can be set for the module internal clock.
After setting or resetting the date and time, use F1
“Set Time” to store.
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Operating Instructions
Setup Functions
Objects
The various module Firmware Object classes can be selected and the functions
most used by the user are directly accessible.
For detailed listing see “LC Injector HTC/HTS-xt System Flow Chart based on
PAL Firmware version 4.1.X (HTC)” on page 246.
44
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
3
Description and Installation
General System Overview 47
Specifications 48
Performance Specifications 49
Electrical Specifications 50
Physical Specifications 51
Operating and Environmental Requirements
Sound Pressure Level 52
Software Requirements 53
Regulatory Compliance Requirements 53
Installation 54
Unpacking the Components 54
Assembling the LC Injector HTC/HTS
Electrical Connections 86
52
55
Object Positions 88
Defining Object Reference Positions 88
Description of Object Reference Positions
91
Injection Valve 105
Valve Drives and Valves – General Remarks
Injection Valve Flow Path 106
Valve Needle Guide 112
105
Interfacing the LC Injector HTC/HTS to other Devices
Synchronization and Output Signals 115
PAL Software Program 118
PAL Loader Program 118
PAL Object Manager Software
115
119
Establishing Communication with Computer
COM Port Settings 120
Setting up LAN Communication 122
120
Agilent Technologies
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3
Description and Installation
Setup Functions
Special Functions 126
How to Access “Extended User Mode”
Section “F3-Setup” 127
Section “F3-Setup” / Objects 134
126
Troubleshooting 146
No or very low detector signal is observed. 146
Mobile phase and/or sample flows out of the wrong ports on LC
injection valve. 147
Sample is backing up on the valve needle guide. 147
Syringe does not fill properly. 148
Sample peaks or responses are not reproducible. 148
Peak distortion or tailing. 149
Excessive carryover between samples 150
Unexplained collisions of the Injection Unit. 151
“Object Not Detected” (Object e.g. Tray1, LCVlv1, Stk1- 01) 152
“Object Collision before Z Tolerance” (“Object” e.g. Tray1, LCVlv1,
Stk1-01) 152
“Invalid Motor Status Motor AUX1”. (AUX2) 152
Special Considerations for Troubleshooting 153
Standard Chromatographic Tests 163
Replacing Parts 170
Warnings and Cautions 170
Replacing Control Board 171
Injection Valve / Valve Rotor 173
Injection Unit 174
DLW Pump Module 174
DLW Wash Station 174
DLW Syringe 175
DLW Syringe Holder Assembly 175
Replacing the DLW-2 Holding Loop 176
DLW Option Spare Parts 178
Maintaining the LC Injector HTC/HTS System
189
This chapter gives an overview and information about the installation of your
Agilent 1290 Infinity LC Injectors HTC/HTS.
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Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Description and Installation
General System Overview
3
General System Overview
NOTE
Agilent reserves the right to make improvements and/or changes to the product
specifications without notice.
The LC Injector HTC/HTS includes the following hardware:
1
X-, Y-axes assembly;
1
Injection Unit, Z-axis;
1
Dynamic Load an Wash Option;
1
1200 bar Injection Valve
1
Keypad terminal;
1
Safety Guard;
1
Standard standalone supports, 215 mm.
Optional TraySets (Tray Holder and Tray) and/or Stacks of various types and
in different variations are required and can be added.
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Description and Installation
General System Overview
Specifications
Table 21
48
Specifications
Type
Specification
System
XYZ robot with syringe-only concept, no tubing in sample path
Sample capacity
HTC Sample capacity
Up to 24 shallow well-plates (96 or 384 )
Up to 12 deep well-plates (96 or 384 ), or 648 2 mL vials
Up to 12 shallow well-plates (96 or 384 ), thermostatted
Up to 6 deep well-plates (96 or 384 ), or 324 2 mL vials, thermostatted
HTS Sample capacity
Up to 24 shallow well-plates (96 or 384 )
Up to 18 deep well-plates (96 or 384 ), or 972 2 mL vials
Up to 12 deep well-plates (96 or 384 ), or 648 2 mL vials, thermostatted
Syringe Size
100 µL
Loop sizes for DLW
and maximum
pressure rating
2 µL (standard) 1200 bar max., 5 µL, 10 µL, 20 µL1000 bar max., 50 µL and
100 µL600 bar max.
Minimum Sample
Volume
1 µL from a 3 µL sample in 1 mL tapered micro vial in liquid injection mode
Injection volumes
Up to 100 µL with standard setup, up to 5 mL with additional hardware
Injection precision
<0.5 % with 100 µL syringe and 2 µL loop, overfill 7 µL
LC Injector
1 electrically actuated fast switching 6-port valve from Rheodyne (1200 bar)
Needle, syringe valve
cleaning
Dynamic load and wash (DLW) principle: two different solvents
Other wash options are available
Carryover
Typically < 0.004 % with DLW Method: Chlorohexidine, 0.6 mg/L dissolved
in H2O : ACN = 90 :10 , 0.1 % TFA Blank Solution: H2O : ACN = 90 :10 , 0.1 %
TFA
Local control
Handheld control panel with 4 function keys, graphical LCD display
Electronic control
•
•
•
•
Thermal control
4 – 40 °C (settable)
2
3
1
2
RS232C ports
TTL inputs
opto-coupler input
relay outputs
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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Description and Installation
General System Overview
Table 22
Specifications DLW-2 Option
Type
Specifications
DLW Pumps
Two pcs. solenoid diaphragm pump, self-priming - Flow rate range:
100 mL/min at atmospheric pressure (Pump itself, no connections to
HPLC system) - Wetted parts Pumps: Ryton PPS and Kalrez (FFPM)
Transfer Tubing Kit
Two pcs. 1/8" PFA tubes including connection fittings
Solvent Reservoir
1000 mL borosilicate glass including Glass Filter, 40 µm pore size
DLW Wash Station
Wash Station with two solvent ports and one Waste port. Two additional
horizontal Waste inlet ports in front to be combined with Valve waste port.
• Wetted parts:
• Wash Station block: PVDF
• Inserts: sst 1.4404
• Connections at bottom of inserts: PEEK
• Waste Tube: Polyethylene (PE)
DLW Syringe
Assembly
•
Syringe:
Gastight Syringe 100 µL with Valflon tip
• Wetted parts:
•
•
•
•
Glass, PTFE and Valflon
Syringe Needle:
• Gauge 22 (OD 0.72 mm/ID 0.41 mm) Length: 0.51 mm, PTFE Seal
DLW Actuator/ 2/2-way flipper solenoid valve
• Solenoid:
• Wetted parts:
• Body: PEEK
• Seal material: FFKM (Simriz)
DLW Manifold Solvent selector module and Syringe adapter assembly
• Wetted parts:
• Block: sst 1.4435
• Perfluor (O-ring)
Holding Loop:
Stainless steel Tubing, OD 0.72 mm, ID 0.41 mm(Gauge 22) Length
85.5 cm; Loop Volume: 118 µL Internal surface acid passivated.
Performance Specifications
The performance specifications are available as part of a specific, separate
document.
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Description and Installation
General System Overview
Electrical Specifications
Table 23
Electrical Specifications
Parameter
Requirement
Protection Class1
Class I
Over Voltage Category2
Category II
Pollution Degree3
2
Moisture Protection4
Normal (IPXO)
Voltage
36 VDC
Current
3.2 A
Fuse
T6.3 A/250 V
System Power Supply
50
Input line voltage
Grounded AC, 100 to 240 V
Input line frequency
50 /60 Hz
Input power
4A
Output voltage
36 VDC
Output current
4.16 A
1
Protection class describes the insulating scheme used in the instrument to protect the user from
electrical shock. Class I identifies a single level of insulation between live parts (wires) and exposed conductive parts (metal panels), in which the exposed conductive parts are connected to a
grounding system. In turn this grounding system is connected to the third pin (ground pin) on the
electrical power plug.
2
Over Voltage category II pertains to instruments that receive their electrical power from a local level such as an electrical wall outlet.
3
This is a measure of pollution on electrical circuits that may produce a reduction of the dielectric
strength or surface resistivity. Degree 2 refers to normally only non-conductive pollution. Occasionally, however, a temporary conductivity caused by condensation must be expected.
4
Normal (IPXO) – IPXO means that there is NO Ingress Protection against any type of dripping or
sprayed water. The X is a place holder to identify protection against dust if applicable.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Description and Installation
General System Overview
3
Physical Specifications
Table 24
Physical Specifications
Parameter
Requirements
LC Injector HTC
Height
648 mm (25.5 in)
Depth
385 mm (15.2 in)
Width
545 mm (21.5 in)
Weight
8 kg (18 lbs.) without accessories
LC Injector HTS
Height
648 mm (25.5 in)
Depth
385 mm (15.2 in)
Width
828 mm (32.6 in)
Weight
10 kg (22 lbs.) without accessories
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Description and Installation
General System Overview
Operating and Environmental Requirements
Table 25
Operating and Environmental Requirements
Parameter
Requirements
Operating Temperature Range
4 to 40 °C (39 to 104 °F)
Maximum Relative Humidity
75 %, non-condensing
Bench Space
At least 24 cm (10 in.) at the rear.
Access to power switch(es) and power cord(s).
Clean, level and smooth surface.
Solid bench plate.
Vibration
Negligible
Static electricity
Negligible
Sound Pressure Level
Table 26
Parameter
Requirements
Sound Pressure Level
Measured value: 62 dBA1
(PAL System used for measurement)
One meter from the equipment in the direction of
maximum sound pressure level. According to UL
610107A-1, 1st edition, clause 12.5. Limit <
85 dBA
1
52
Sound Pressure Level
dBA = ”A weighted” sound pressure level
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Description and Installation
General System Overview
3
Software Requirements
The various PAL Software programs, such as PAL Loader or PAL Object
Manager, are operated with Microsoft Windows Operating systems, such as
Windows XP and Vista.
The Agilent 1290 Infinity LC System can be controlled using PAL control
software, or by Agilent LC/LCMS ChemStation, Masshunter for LCMS and
EZChrom.
Regulatory Compliance Requirements
As of the date of publication, this product is compliant with current RoHS and
WEEE regulations.
• Directive 2002/95EC, RoHS
• Directive 2002/96/EC, WEEE
NOTE
Agilent Technologies reserves the right to make improvements and/or changes to the
product specifications without notice.
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Description and Installation
Installation
Installation
Unpacking the Components
An autosampler system is shipped in several boxes. One box contains the
X-,Y-axes assembly, the Injection Unit, the Keypad Terminal, standalone
Supports, connecting cables, Power Supply, 1200 bar Injection Valve including
valve drive, Safety Guard and miscellaneous parts. The Dynamic Load and
Wash accessories and all optional parts, such as TraySet, Stack Cooler, second
valve drive assembly, etc., are shipped in separate boxes.
1 Open the box and first remove the accessory boxes and the Injection Unit
before attempting to remove the X-, Y-axes assembly.
2 Carefully lift the X-,Y-axes assembly and remove it from the box. Hold the
Y-axis in place while the assembly is being removed from the box. Set the
X-,Y-axes assembly on a bench.
3 Unpack the remaining small boxes and any other accessories.
4 When placing the LC Injector HTC/HTS onto a stable surface, make sure
that no objects interfere with either the Y-axis or the Injection Unit
throughout the entire range of potential movement.
54
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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Description and Installation
Installation
Assembling the LC Injector HTC/HTS
Before beginning the assembly process, determine approximately where
the LC injection valve and keypad terminal will be located. The terminal
may be mounted on either side of the X-axis.
1 If a Stack (a Tray Holder with multiple drawers) was shipped with the LC
Injector HTC/HTS, loosen the Torx screws on the two mounting clamps
located on top of the stack.
2 Move the X/Y-Carriage to the center of the X-axis and temporarily place the
X-axis assembly on top of the Stack with the mounting clamp teeth fitting
into the grooves on the bottom of the X-axis.
NOTE
For Systems without a Stack, support the X/Y-Carriage in a suitable manner (a sturdy
cardboard box can be used) before attempting to install the legs.
3 Install the legs near the ends of the X-axis. Loosen the Torx screws on the
mounting clamps and then fit one leg into the grooves in the X-axis. Be sure
that the clamps fit completely into the grooves. Tighten the Torx screws
until the legs are firmly in place.
4 Double check whether the leg claws are correctly attached to the X-axis (see
Figure 8 on page 55).
Lgdc\
Figure 8
8dggZXi
Attachment of Mounting Claws
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3
Description and Installation
Installation
Installing the Injection Unit
Figure 9
NOTE
Attaching the Injection Unit (shown freestanding for clarity)
Installation of the Injection Unit should be done carefully. When installing it for the first
time, have someone hold it in place while the mounting screws are inserted.
(See Figure 11 on page 58).
56
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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Description and Installation
Installation
1 Remove the three Torx mounting screws A, B, and C, used to fix the
Injection Unit to the Y-axis
2 Connect the ribbon cable (see Figure 10 on page 57).
G^WWdcXVWaZXdccZXi^dc
<j^YZ]daZh
6
7
<j^YZe^ch
Figure 10
Connecting the Injection Unit Ribbon Cable
3 Hold the Injection Unit in place against the Y-axis. Make sure the two
locating pins on the Y-axis fit into the two guide pin holes on the Injection
Unit.
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Description and Installation
Installation
4 Place one of the screws onto the end of the supplied Torx driver. Slide the
clear plastic cover on the Injection Unit all the way to the top. Locate the
three large holes in the black anodized frame attached to the Z-axis inside
the Injection Unit. Slide the frame upwards until the top hole is centered on
the top threaded hole at the end of the Y-axis. Insert and securely tighten
the Torx screw A (see Figure 11 on page 58).
Figure 11
58
Inserting the Injection Unit Mounting Torx Screws
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
3
Description and Installation
Installation
5 Install the two remaining Torx screws B, C in the left and right mounting
holes, respectively. It may be necessary to move the elastic cord slightly to
the left to insert the Torx screw C into the right-hand hole.
Figure 12
LC Injector HTC/HTS with Injection Unit and Standalone Supports
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Description and Installation
Installation
Installing the Keypad Terminal and Safety Shield
Figure 13
Installing Keypad Terminal
1 Install the safety shield on the left and right sides to the outside of the Xaxis. Use the provided, longer thumbscrew on the side where you plan to
install the keypad.
2 Install the Keypad mounting bracket on either the right or left side of the
X-axis.
3 Connect one end of the white coiled cable (Part No. SS8J-700) to the Keypad
and the other end to the TERMINAL (SER3) interface jack on the rear side
of the X-axis. For details see Figure 33 on page 86 or Figure 34 on page 87.
4 Place the Keypad terminal onto its mounting bracket.
NOTE
Do not interchange the Terminal with LAN cable connectors.
Installing the Power Supply
1 Locate the power supply, the DC power cable, and the AC power cable.
2 Set the power supply switch to the OFF position.
3 Connect one end of the DC power cable to the power supply and the other
end to the POWER connector at the rear side of the X-axis.
4 Connect the female end of the AC power cable to the power supply. Then
connect the male end to an AC power outlet.
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Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Description and Installation
Installation
3
Installing the LC Injection Valve
1 Locate the blue, valve drive. It will have one clamp that is identical to other
object clamps. Loosen the clamp as above (see Figure 14 on page 61).
Figure 14
Installing the Injection Valve Drive
2 Attach the valve drive to the X-axis and tighten the mounting screw.
NOTE
To avoid delay volume between sample injection point and detection, the injection valve
should be located near the detection device.
3 Connect the control cable from the valve drive to AUX1.
NOTE
The installation of another type or multiple Valve Drive(-s) is described in “Valve Drives and
Valves – General Remarks” on page 105.
4 The injection valve and rotor are pre-installed on the valve drive.
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Installation of the DLW Option
General System Overview
Figure 15
DLW-2 Option Components
DLW: Dynamic Load and Wash
The DLW Option represents a new concept in wash stations, combining an
injection cycle with wash steps. The combination of both injection and wash
steps – usually separate procedures – minimizes cycle time and carry-over.
The two characteristic features of the DLW Option are:
• The sample solution never comes in contact with the syringe itself; it is held
‘sandwiched’ in a so-called Holding Loop.
• Wash solvents are pumped from the rear to the front of the DLW system, to
intensely purge all critical surfaces which have come in contact with the
sample.
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The DLW option consists of two self-priming micro pumps (mounted on a
dedicated bracket), the wetted parts are composed of Ryton PPS and Kalrez
(FFPM). The ‘IN’ ports are connected to the wash solvent bottles and the ‘OUT’
ports are connected to the DLW manifold, which is part of the dedicated DLW
Syringe Holder assembly. A ‘Holding Loop’ separates the syringe and the DLW
actuator to avoid the sample coming in contact with these parts.
TThe syringe and Holding Loop are preloaded with Wash Solvent #1 at the
start. The sample is picked up and remains separated from Wash Solvent #1 by
an air gap. After loading the loop and injection, Wash Solvent #1 is pushed
into the system, followed directly by Wash Solvent #2 to flush the critical valve
paths.
The DLW Syringe Assembly is then moved to the wash station for further
cleaning steps and to prepare the Syringe and Holding Loop for the next cycle.
Further details are given in “DLW Cycle Step-by-Step” on page 211.
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Development of the PAL DLW Option
The DLW Option has undergone several development steps. The current
version, DLW-2 Option, contains several improvements over the earlier
versions:
• Holding Loop
The DLW-2 Holding Loop consists of one single piece from the needle to the
loop. This eliminates several tube connections, the replacement needle and
the Flow diverter. The loop is made of high quality stainless steel and the
inner surface is passivated with acid.
• DLW Syringe Holder Back-plate
The DLW-2 Syringe Holder back-plate has changed in size and form to allow
a smoother replacement of the holding loop. The Actuator is supported
from the backside to provide higher mechanical stability.
• DLW Pump Assembly
The DLW-2 Pump assembly is newly equipped with a housing to protect the
electrical parts from solvent splashes.
NOTE
64
This Manual covers the DLW-2 Option. The technique as such has been established with
previous versions and is called the ‘DLW technique’. Most of the parts are used for the
‘DLW Option’ as well as for the ‘DLW-2 Option’. DLW-2 specific parts or their usage are
emphasized. Otherwise, the general term ‘DLW’ is used which applies to the earlier
versions and the current version ‘DLW-2’.
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Hardware Requirements
The DLW option can be used with Agilent 1290 Infinity LC injectors in
combination with the Rheodyne ultra high pressure valves.
Software Requirements
The DLW option can only be operated with Firmware 1.2 or higher. It can be
controlled using PAL Control software, the Cycle Composer or any CDS
(chromatography data system) software that controls the PAL System
including those using the Cycle Editor for PAL ICC interpretation.
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Unpacking the Components
The DLW-2 option is shipped in one box. Check for the following items:
Figure 16
66
DLW-2 Option Components
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Item p/n
Description
1
G4277-60510
DLW Pump Module
(re-order for one solvent channel)
2
5190-3259
DLW Syringe
3
G4277-60509
DLW Plunger Holder
4
G4277-60501
DLW Syringe Holder Assembly
5
CD ROM
6
G4277-60513
Wash Station cable to System
7
G4277-60512
DLW Tubing Kit
8
G4277-60602
DLW Holding Loop (installed in the DLW Syringe Assembly)
9
G4277-60515
Needle Guide Length Tool
10
G4277-60514
DLW Wash Station incl. Waste Tube
11
G6500-88035
Solvent Bottle Transfer Line including Glass Filter Solvent Inlet, 40 µm pore
size
12
(2x)
1 L Solvent reservoir bottles
Please refer to Figure 15 on page 62 for an overview of how the various DLW
components are mounted on the system.
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Mounting the Pump Holder
The pump holder should always be mounted on the far left of the X-axis. The
solvent lines are connected to the pump holder. Moving across the pump
holder with the Z-axis could kink the stabilizing wire. To install the holder, it is
recommended to keep the DLW syringe module out of Z-axis.
1 Locate the pump holder and the two supplied Torx screws.
2 Attach the holder to the X-axis as shown in Figure 17 on page 68.
Figure 17
68
Attaching Pump Holder Bracket to X-Axis
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3 The Solvent line assembly is not preinstalled at the factory. The correct
mounting is shown in Figure 18 on page 69. Select the end of the guide wire
with the longer tubing ends and connect the guide wire with the screw M4 x
8 and the corresponding serrated lock washer to the back side of the
module. Note the guide groove, which orients the wire.
NOTE
Always install the DLW Tubing (from the kit) to the pump module first. The other end,
connected to the DLW Syringe Assembly, should be connected while the assembly is not
yet installed onto the injection unit.
Figure 18
Installing the Solvent Tube Assembly to the Pump Holder
4 Connect the solvent tubing of the DLW Tubing kit to the pump module,
using the premounted nut. Maintain this order: Solvent 1 connected to the
left, and Solvent 2 to the right of the pump module, when viewed from the
front of the system. This avoids any confusion when refilling or changing
solvents.
5 Connect the solvent lines from the solvent reservoir bottle to the “Inlet”
connectors of the pump module.
6 The electrical connections from the pumps to the PCB sockets, mounted on
the bracket, are done at the factory.
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7 Insert the cable provided into the middle socket, between the other two
sockets on the PCB. For details see Figure 19 on page 70.
Figure 19
Electrical Connections at the Pump Holder Bracket
8 Connect the other end of the cable to the “Was Station” connector on the
Control board, as shown in Figure 20 on page 70.
Figure 20
70
Electrical Connection from DLW Pump Holder to the LC Injector HTC/HTS
Board
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Installing the DLW Wash Station
The DLW Wash Station can be connected anywhere along the X-axis; however,
it is strongly recommended to install it as close as possible to the Injection
Valve. The travel path of the Injection Unit from the valve to the Wash Station
should be as short as possible to keep the cycle time as brief as possible.
Figure 21 on page 71 shows two configurations for installing DLW modules,
favoring a short connection from the injection valve to the other HPLC system
components on the one hand, and always having the DLW Wash station close
to the injection valve on the other. It is important that the DLW Pump Module,
with the connected DLW tubing from the kit, always be attached on the far left
side of the X-axis. Such a configuration also has the advantage that the waste
tube from the injection valve can be connected to the front “Waste Inlet” of the
Wash Station module. This waste tube is part of the DLW Option Kit.
Figure 21
Mounting the DLW-2 Option Modules in combination with the Injection Valve
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1 Locate the DLW Wash Station module.
2 Attach the holder to the X-axis at selected position as shown in Figure 21
on page 71.
3 Connect the Waste Tube from the Injection Valve to the DLW Wash Station.
4 Connect the Waste Tube to the Wash Station Waste Adapter.
5 Replace the dummy plug(s) on the front if used to connect the waste line
coming from the injection valve.
This sequence of waste lines simplifies the system, avoiding several waste
lines running to the waste container.
NOTE
The other two dummy plugs underneath of the waste block are used for the DLW-2 option.
The DLW tubing from the DLW Pump outlet could be connected at these wash station inlet
ports. The functionality of the Fast Wash Station would then be configured using this order
of connections.
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Figure 22
NOTE
3
Attaching the DLW Wash Station to the X-Axis
The waste tube must be positioned below the injection valve. Make sure that the waste
liquid flows into the waste container without any restriction. The waste tube must always
be placed above the liquid level in the waste container.
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Installing the DLW Syringe Holder Assembly
The DLW Syringe Holder differs from standard liquid syringe holders because
the DLW Manifold is attached to the syringe holder. The DLW
Actuator/Solenoid and the Holding Loop connecting the syringe inlet (bottom)
and Syringe Needle are attached to the DLW Manifold. All of these parts are
preinstalled at the factory. See Figure 23 on page 74.
Figure 23
74
Front View of the DLW Syringe Holder Assembly
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CAUTION
Needle damage
➔ Before a syringe is inserted, the position 'Change Syr' should first be verified. The
position for 'Change Syringe' should not occur above an object where the needle
could collide with a vial.
➔ Path:
➔ Menu > Utilitites > Syringe 'F3' 'Change Pos'.
➔ Move the Injection Unit to a location where no collision can occur with the Z-axis by
selecting the X- and Y- axes appropriately. The value for the Z-axis is given as a
default and a change of this position is not necessary in standard operation.
➔ This precaution helps avoid needle damage during routine operation. Nevertheless,
the description below on how to install the DLW Syringe Assembly recommends
switching off the unit and inserting the assembly without using the command
'Change Syringe'.
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Preparing and Installing the DLW Syringe
1 Prepare the DLW Syringe by inserting the DLW Plunger Holder.
a First, move the plunger manually down to the stop position.
b Release a slight amount of pressure from the plunger tip by pulling a
fraction of a mm backwards.
c Install the DLW Plunger Holder and tighten the Allen screw (Allen Key
#6) firmly.
Figure 24
NOTE
76
Inserting DLW Plunger Holder on the DLW Syringe
It is critical that syringes are primed before beginning sample preparation. Prime every
liquid syringe first manually before inserting into the system.
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2 Screw the prepared DLW syringe into the holder. Hold the syringe at the
lower metal mount while tightening the syringe.
Figure 25
NOTE
Inserting the DLW Syringe into the Holder
Holding the glass barrel while tightening can damage the seal where the glass meets the
metal.
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Connecting the DLW Tubing to the Syringe Holder
NOTE
It is recommended to connect the DLW Tubing from the kit to the Syringe Holder while the
assembly is not yet inserted in the X-axis.
NOTE
The wetted parts of the DLW Actuator/Solenoid are composed of PEEK for the body and
FFKM (Simriz) for the seal material. PEEK exhibits excellent chemical resistance to most
commonly used chemicals. However, the following solvents are not recommended for use
with PEEK: DMSO, THF, methylene chloride (dichloromethane), nitric acid, sulfuric acid. For
more details, see the compatibility tables provided by the manufacturer of PEEK materials
or components.
1 As described under “Mounting the Pump Holder” on page 68 the end of the
the DLW kit sleeved tubing with the short tubes has to be connected to the
DLW Syringe Holder Assembly.
2 Use the groove in the block at the back side of the DLW Manifold to orient
the guide wire. Use the M3x5 screw provided and the corresponding
serrated lock washer to connect.
3 Test if the wire tension is enough to keep the tubing in an upright position
but low enough to move the syringe holder manually along the X-axis.
NOTE
The design of the DLW Manifold and the concept of washing by active pumping and closing
of the lines by the DLW Actuator theoretically does not require a predefined solvent line
position, in theory. Problems could occur if solvent is refilled or exchanged while the tubing
still contains solvent from the previous setup. Always keep the lines at the same position,
and careful prime the entire system to prevent unnecessary confusion. See also “DLW
Principles and Operating Details” on page 200.
4 Connect solvent line 1 to the upper port and solvent line 2 to the lower port
at the left side of the DLW Manifold.
78
NOTE
The order of the tube connections, upper and lower positions should be consistent.
NOTE
For certain applications it is crucial not to mix types of solvents, for example biofluid sample
solutions should not come in contact with highly concentrated organic solvents.
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Inserting the DLW Syringe Holder Assembly into the Injection Unit
NOTE
It is recommended to insert the DLW Syringe Holder assembly when the LC Injector
HTC/HTS is powered off.
1 Move the Injection Unit aside manually to allow free movement of the
syringe slider. Lower the syringe slider as shown in Figure 26 on page 79 in
order to gain space for installing the DLW Syringe assembly.
NOTE
Match the magnetic pins of the syringe holder using the counter positions on the syringe
slider.
The position of the Guide Pin Holes can be matched with the ‘Positioning DLW Guide Line’
which is marked on the DLW syringe plate.
Figure 26
Inserting the DLW Syringe Holder Assembly into the Injection Unit
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2 Press the syringe holder firmly against the Z-axis slider to ensure that the
holder clicks into place.
HINT
Do not press against the DLW Actuator/Solenoid. Its mounting to the DLW Manifold is
fragile.
3 Tighten the knurled screw to fix the Holder Assembly to the syringe slider.
(See Figure 27 on page 80.)
Figure 27
80
Fixing the Syringe Holder to the Syringe Slider
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4 Move the plunger up (plunger holder) until the thread of the screw catches
the thread of the plunger bushing. Tighten the screw to fix the plunger
holder. (See Figure 28 on page 81.)
Figure 28
Connecting the DLW Syringe Plunger Holder
5 Tighten the holding screw to secure the syringe holder position.
6 Move the lower needle guide carefully up and down to make sure that the
needle tip does not catch on the guide.
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Injection Valve Plumbing and Connection to DLW Wash Station
1 Connect the injection valve Waste Line from port 6 for the Agilent 1200 bar
LC injection valve 5067-4123, as shown in Figure 29 on page 82, to the front
Waste Port of the Wash Station
NOTE
If two injection valves are used in the LC Injector HTC/HTS configuration, the second
waste line can be connected to the lower front waste port of the DLW Wash Station. If only
one connector is used, apply a dummy plug to the second port on the Wash Station.
2 Place the Waste Tube from the DLW Wash Station into the waste container.
NOTE
The waste container must be positioned below the injection valve. Make sure that the
waste liquid flows into the waste container without restriction.
Figure 29
NOTE
82
Solvent Line Plumbing on Rheodyne Valve PD Series
The Agilent specific Rheodyne Valve with Agilent PNo. 5067-4123 is connected in the same
way to the HPLC system as shown in “Installation” on page 108. For details see the specific
valve description.
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Electrical Connections for the Single Injection Valve Setup
NOTE
Always switch the LC Injector HTC/HTS power supply OFF before connecting or
disconnecting the LC Injector HTC/HTS DLW cable or any other LC Injector HTC/HTS
accessory cables!
The electrical connection of the DLW option is identical for the PAL HTS
and the HTC models. The same board is used for both models.
1 The schematic in Figure 30 on page 83 shows an installation with the valve
drive connected to the 'AUX' interface. If a serial or multiposition valve
drive is used, see the schematics for 2- or 4-injection valve combinations.
Figure 30
Electrical Connection, DLW Option on a PAL
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Installation
Installing a Microplate Stack
1 If a Stack (a Tray Holder with multiple drawers) was shipped with the LC
Injector HTC/HTS, loosen the two Torx screws on the two mounting clamps
located on top of the Stack. The example below shows the two stack models
with 3 and 6 drawers.
Figure 31
NOTE
Installing a Microplate Stack with 3 or 6 Drawers
Installation of Stack and Tray Cooler is described in detail in the Addendum to the User
Manual “MT/DW Tray, Stack, and Cooler Upgrades for Agilent LC Injectors HTC/HTS”.
2 Carefully lift the X-axis assembly on top of the Stack with the mounting
clamp teeth fitting into the grooves on the bottom of the X-axis.
3 Be sure that the clamps fit completely into the grooves. Alternately tighten
the two Torx screws until the two mounting clamps are firmly in place.
4 Double check whether the Stack clamps are correctly attached to the X-axis
(see Figure 8 on page 55).
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Installing a Trayholder
1 If a Tray Holder was shipped with the LC Injector HTC/HTS, loosen the two
Torx screws on the two mounting clamps located on top of the Tray Holder
legs.
Figure 32
Installing a Tray Holder
2 Install the Tray Holder with the mounting clamp teeth fitting into the
grooves on the bottom of the X-axis.
3 Be sure that the clamps fit completely into the grooves. Alternately tighten
the two Torx screws until the two mounting clamps are firmly in place.
4 Double check whether the two Tray Holder clamps are correctly attached to
the X-axis (see Figure 8 on page 55).
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Electrical Connections
NOTE
Always switch OFF the power supply before connecting or disconnecting any accessories
cable!
NOTE
Do not interchange the Terminal with LAN cable connectors.
1 Before defining the object positions, make sure the LC Injection Valve, Fast
Wash Station and Keypad Terminal are correctly connected to the X-axis
rear side (see Figure 33 on page 86 or Figure 34 on page 87).
Figure 33
86
Electrical Connections - LC Injectors HTC
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Figure 34
3
Electrical Connections - LC Injector HTS
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Object Positions
Defining Object Reference Positions
The objective is to define the Reference Positions for all autosampler Objects.
Make sure the Tray Holder(s), Valve Drive(-s)and Wash Station are properly
mounted to the autosampler X-axis. The following description is an example of
how to teach the reference position for a Tray Holder. The described
procedure is common to all autosampler Objects.
Figure 35
NOTE
88
Object Reference Position
Remove the Syringe Adapter from the Injection Unit before performing the following steps.
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For a Tray Holder, the Reference Positionis a hole (slightly larger than the
lower needle guide) in the base plate of the holder. The lower needle guide
should be centered in the hole with the bottom of the needle guide flush
with the bottom of the base plate (see Figure 35 on page 88).
1 Switch ON the autosampler power supply.
2 Observe the keypad display. The model name will display along with the
software version number. The Job Queue menu screen will then display.
3 When the "Job Queue" menu displays, complete the following sequence
(common to all objects):
BZcj
HZije
DW_ZXih
IgVn=daYZgh
CVbZYIgVn=daYZg
Figure 36
1
Menu Screen Object Trayholder
"Named Tray Holder" represents a predefined Tray Holder, (e.g. THldr1 or Stack1).
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4 After selecting "Named Tray Holder" the X- Y- Z-positions for the object will be
displayed (see Figure 36 on page 89).
5 Highlight item Position X with the cursor bar and press ENTER. The Injection
Unit will move to the previously defined X-axis position.
6 Rotate the outer knob to adjust the X-axis position to the Tray Holder
reference position.
7 Press the inner knob to ENTER the Position X value.
8 Repeat steps 5-7 for Position Y and Position Z.
9 If F3 "Moveto Zero" is activated, the Injection Unit will move to the HOME
position;
10 Verify the defined X-, Y-, Z-positions by pressing F1 "Check Pos".
Figure 37 on page 90 illustrates possibilities of correction. Details for teaching
are given below.
Figure 37
90
Inclined Tray, Corrections for X-, Y-, Z-Axes
NOTE
The release of Firmware version 4.1.X makes possible the correction of all three axes, X-,Y-,
Z, in the firmware class “ Tray.” The correction of the inclination can be made for a tilted
Tray in the direction of the row and/or the column. Teaching is possible in the ” Utilities”
section by using the path: Menu/Utilities/Trays.
NOTE
These functions are not available for the autosampler if operated solely on PAL Firmware
version 2.4.X or higher, but neither with firmware version 3.X.X. nor with version 4.X.X.
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Description of Object Reference Positions
Trayholder (e.g. THldr1)
Figure 38
Tray Holder Reference Position
For Tray Holder, the Reference Position is a hole in the base plate of the
Holder. The lower needle guide should be centered in the hole with the bottom
of the needle guide flush with the bottom of the base plate.
Figure 38 on page 91 shows the example of a Tray Holder for a single Tray (e.g.
“THldr1” for “Tray 1”) and a Tray Holder for four Trays (e.g. “Tray 1” with Tray
Type “VT54”, “DW96” or “MT96”, etc.).
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Tray Holders Stack or Stack Cooler with 3 or 6 Drawers
Figure 39
Stack Reference Position
The Reference Position for a Stack or Stack Cooler is identical. The reference
position in the case of a 3-Drawer Stack is a hole in the upper drawer and in a
6-Drawer Stack in the second drawer from the top (same height level), as
shown in Figure 39 on page 92.
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Trays
In general it is not necessary to teach a Tray position. The Tray Holder has a
teaching position, as described above. A Tray is assigned to the Tray Holder.
The numbers (values) for the relevant items are stored in the firmware object
“Tray”. To complete the combination of a Tray Holder and a Tray, it is
necessary to assign the corresponding “Tray Type” to the “Tray.” The “Tray
Type” contains the geometric data for the Tray and information, such as how
many samples are in a row and a column in the Tray. A Tray Type defines the
pattern and sampling sequence of sample location within a Tray. For details,
see below.
As described above, the Tray is physically placed in the Tray Holder, the
firmware assigns the Tray to the Tray Holder and the Tray Type to the Tray.
This is routine procedure as long as no special circumstances need be
considered, such as customized Trays or Tray Types.
Autosampler Firmware version 4.1.X now makes a correction possible if the
Tray surface is not exactly horizontal and planar but is inclined in one or more
axes. At version 4.1.X this is standard and a dialog window pops up the
moment the Tray has been checked-out at the corners to verify vial positions
and heights. How to correct a possible inclination is explained below.
Figure 40
Demonstrating a possible Inclination of Tray in X-, Y-, Z-Axes.
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Object Positions
Definition of a Tray Row and Column
The autosampler system defines Rows and Columns by the order in which
samples are treated. A “Row” is not associated with an X- or Y-axis. The
definition is visualized below with Tray Type VT32 and DW96:
Gdl
8dajbc
8dajbc
Gdl
Figure 41
94
Tray Type VT32 and Tray Type DW96
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Correction for Sample Position
The following description of the correction is done by using the path to the
“Utilities” section.
After selecting the Utilities function “Trays”, verify whether the Tray Type
assigned corresponds to the Tray placed into the Tray Holder. If this is
verified, the vial positions can be checked at the three corners of the Tray.
This is activated by the “F3” function key.
BZcj
Ji^a^i^Zh
IgVn
HZaZXiZYIgVn
HZaZXi;jcXi^dc@Zn;(
BdkZid%%&
A dialog window opens, displaying a note to “Adjust offset X-, Y-, Z for Position
001”. Continuing in this dialog will provide the user the opportunity to adjust
all three axes perfectly by the top of the vial. Please note that this step is only
valid for position number 001.
Figure 42
Adjusting “Offset X,Y,Z” for Tray Position 001
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To check the next corner of the Tray, activate function key “F3” again, example
Tray Type “VT32”, position 008.
The next information provided by the pop-up window asks to correct a
possible inclination of the “Row”. Continuing the dialog provides the user the
opportunity to adjust the three axes perfectly to match the vial top – or in case
of a well-plate, the top of the plate.
Figure 43
Adjusting Row Inclination at second Tray corner position
The last step will be to move to the next corner of the Tray by again activating
function key “F3”. This will be position 32 in the example of Tray Type “VT32”.
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A possible inclination of the Tray column can be corrected in the same manner
as described above.
Figure 44
Adjusting Column Inclination at Third Tray Corner Position
The three corner points are now adjusted for a possible inclination of the Tray
in any axis. A possible deviation from an ideal axis position for the other vials,
caused by variance from the horizontal, is interpolated by the system without
further teaching.
NOTE
For routine work using standard vials it is not necessary to compensate for a possible
inclination as long as the vial detection is within the “Z-Tolerance” range.
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Object Positions
Loading the LC Injector HTC/HTS DLW Object
If the version of the PAL Object Manager List installed on the computer is not
'PAL-xt Object Lists Rev. C', copy the provided folder 'Wash Station Option' (See
Figure 45 on page 99.) from the CD-ROM to the Object Lists folder which has
been installed with the Object Manager. This software is usually installed in
the following path:
C:\Program Files\PAL\Object Manager\Object Lists
Using PAL-xt Object List Rev. C or higher, the DLW Option object will become a
part of the list in the folder 'Wash Stations' folder. The following Object List is
then available:
PAL DLW: DLW Option Firmware Object List
NOTE
The instructions in the following section have to be followed for upgrades only. All Agilent
1290 Infinity LC Injector HTC/HTS are configured with the right objetcs from the factory.
1 Open the Object Manager and verify the revision number of the Object Lists.
If PAL-xt Object Lists Revision level 'C' (or higher) is available, continue as
described below.
If a 'PAL Object Lists' version is used (for example Rev. K or L), copy the
provided DLW Object List into the Object Manager folder as described
above.
2 Select the 'PAL DLW' Object list and click 'Send selected Object Lists to PAL'.
3 Close the Object Manager.
The DLW Object list contains the following LC Injector HTC/HTS Firmware
Objects which are loaded with a single command as described above:
98
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Description and Installation
Object Positions
Table 27
3
DLW Object List
Object Class
Object Name
Motor Drive
MPlgMed
Heater
HDLW
Syringe
100ulDLW
Wash Station
Wash1
Wash2
Injector
Waste
PWR Event
Pwr-Out1
Pwr-Out2
Figure 45
NOTE
Object Manager; choosing Object List Folder 'Wash Stations'
For more details on the PAL Object Manager software, see the Addendum to the PAL User
Manual PAL Object Manager Software.
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Description and Installation
Object Positions
Wash Station Reference Points
Wash1/Wash2 Reference Points
For the DLW Wash Station Option, the reference positions for 'Wash1' and
'Wash2' are the two holes on top of the Wash Station assembly. See Figure 46
on page 100.
The lower needle guide should be centered on these holes with the bottom of
the lower needle guide lightly touching the surface of the Wash Station
assembly.
Figure 46
DLW Wash Station Wash1/Wash2 Reference Points
1 Press 'HOME' on the handheld controller and select the sequence Menu >
Setup > Objects > Wash Stations > Wash1.
2 Enter Wash1 and adjust the X-, Y-, Z-positions until the lower needle guide
of the Injection Unit centers on top of the Wash station assembly. Fine tune
the Z-position until the bottom of the lower needle guide is flush with the
top surface.
3 Repeat steps 1 – 3 for Wash2 by using the path: Menu > Setup > Objects >
Wash Stations > Wash2
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Description and Installation
Object Positions
3
Waste Reference Point
The reference position for the Waste Port – a hole (slightly larger than the
needle guide) – has been placed in the middle positions of 'Wash1' and 'Wash2'.
(See Figure 47 on page 101.) The lower needle guide should be centered on
this hole with the bottom of the lower needle guide lightly touching the surface
of the Wash Station assembly.
Figure 47
NOTE
Waste Reference Position
The Waste position is defined as an 'Injector' by the software. It is classified in the Object
category 'Injectors'.
1 Using the handheld controller select the sequence: Menu > Setup > Objects >
Injectors > Waste 1.
2 Enter Waste and adjust the X-, Y-, Z-positions until the lower needle guide of
the Injection Unit centers on top of the Wash Station assembly. Fine tune
the Z-Position until the bottom of the lower needle guide is flush with the
top surface.
3 Press 'F1' to verify the position and 'F4' for 'Home'.
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Description and Installation
Object Positions
Injection Valve Reference Points
For an LC Valve, the reference position is the valve needle guide fitting
mounted on the top valve port (see Figure 48 on page 102).
Figure 48
LC-Valve Reference Point
The lower needle guide of the Injection Unit is centered on the valve needle
guide fitting. Adjust the Z-Position so that the bottom of the lower needle guide
just touches the surface of the valve needle guide fitting. Then reduce this
value by 2.0 mm (see Figure 49 on page 102).
'bb\Ve
Figure 49
NOTE
102
Positioning the Z-Axis Needle Guide on the Valve Needle Guide
To adjust the Valve Needle penetration value, see “Defining the Valve Penetration
Value” on page 103.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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Description and Installation
Object Positions
Defining the Valve Penetration Value
NOTE
Before performing the following steps, make sure that Object positions X, Y, and Z for LC
Vlv1 are properly defined and the complete DLW Syringe assembly, is inserted into the
injection unit.
Complete the following steps to define the Valve Needle Penetration Depth:
1 Using the handheld controller select the sequence: Menu > Setup > Objects >
Injectors > LC Vlv1
2 Press F1 'Check Pos'. The Injection Unit moves to the previously defined LC
Vlv1position.
3 Highlight the item 'Needle Penetr' with the cursor bars by pressing ENTER.
4 Slowly rotate the outer knob to adjust the needle penetration depth.
The syringe moves down stepwise into the injection port.
5 When the syringe needle tip enters the valve needle guide, slow down the
Z-movement again.
NOTE
Always observe the syringe needle during this adjustment step.
6 Move down stepwise until the bottom of the cross bar of the DLW Needle
Holder Assembly is flush with the lower line of the 'DLW Needle Adapter
Block' (see Figure 50 on page 104).
7 Press ENTER to save the needle penetration depth value.
8 Verify the defined Needle Penetration Depth value by repeating steps step 3
on page 103 to step 6 on page 103.
NOTE
If a 'click' sound occurs, this means the ball-spring loaded 'Needle Holder' is completely
compressed. STOP the Z-axis downwards movement immediately. Rotate the outer knob
stepwise in the opposite direction until the gap in the spring loaded “Needle Holder” is
approximately 1 mm, or the bottom of the 'Holder Assembly' is flush with the lower surface
of the 'Needle Adapter Block', as in Figure 50 on page 104. Repeat the needle penetration
test and verify the needle penetration depth.
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3
Description and Installation
Object Positions
Figure 50
Teaching the Needle Penetration Depth Value
An example of the correct and incorrect positioning of the syringe needle tip
in the valve inlet port is shown in Figure 51 on page 104.
Figure 51
Correct and incorrect Valve Needle Penetration Depth
The ball-spring loaded 'Needle Holder' presses the square cut needle tip firmly
against the bottom of the valve body. This ensures a constant seal during the
injection process Figure 50 on page 104.
104
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Description and Installation
Injection Valve
Injection Valve
Valve Drives and Valves – General Remarks
A LC Injector HTC can be equipped with 1 LC Injection valve connected and
controlled via the Auxiliary Interface (AUX). A LC Injector HTS can be
equipped with 1 or 2 LC Injection valves connected and controlled via the
Auxilliary Interface (AUX1, AUX2). If more than one valve has to be
configured, or if an auxiliary Interface is occupied by another module, such as
a Dilutor, then Multi Position or Serial Valve Drives are required.
NOTE
The LC Injector HTC is equipped with the control board “APR Control-xt” which provides
two auxiliary interfaces, “AUX1” and “AUX2”. The choice to connect one more valve drive
is provided.
The standard LC injection valve is a six-port 1200 bar Rheodyne Injection
valve.
The six-port valve is connected to the detection system according to the flow
path shown below. Plumbing diagrams for other available six-port injection
valves are provided below as well.
Details are described in the “Valve Drives and Valves – General Remarks” on
page 105.
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Description and Installation
Injection Valve
Injection Valve Flow Path
The Agilent Injection Valve 5067-4123 is a SupraLife™ 2-position, 6-port Rapid
Replacement Pod equipped with a vertical inlet port, a pinned drive and
designed for very high pressure applications. The concept is based on a flat
plate rotary valve.
Plumbing Diagram
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Figure 52
106
6-Port Agilent Injection Valve 5067-4123
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Description and Installation
Injection Valve
3
Specifications
Table 28
Specifications
Valve Stator:
SPC-5 coated stainless stator
Rotor:
RPC-13 Polyimide based rotor seal
Bore Size:
Vertical Port: 0.25 mm (0.010 in)
Ports 2 to 6: 0.20 mm (0.008 in)
Volume for Vertical Port:
0.75 µL
Ports and Engraving:
Ports 2 to 6: 0.07 µL
Engraving: 0.05 µL (standard engraving)
Valve Volume Full Loop:
Full Loop: 0.87 µL
Partial Loop: 0.82 µL
Connections:
10-32 male threaded fittings, 1/16 in tubing OD
Maximum Pressure:
1200 bar (18000 psi; 120 MPa)
Standard Loop:
2 µL; Loop ID: 0.13 mm (0.005 in)
RoHS:
Compliant
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Description and Installation
Injection Valve
Installation
1 After the mechanical connection of the PAL Valve Drive to the X-axis and
electrical connection to AUX1 at the board, turn on the power of the PAL
System.
The Firmware Object for the Valve type PAL VlvDrv-5067-4123_1 is preloaded,
the Valve Drive will go into the Standby position.
2 Insert the valve body into the shaft until the pin engages in the shaft.
3 Center the valve body by turning the valve body into position until the
vertical port is aligned.
4 Insert the Valve Body completely into the shaft until the pin on top of the
valve engages in the threaded Valve Drive Bracket. Tighten the Valve Body
with the polymer connecting nut.
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Figure 53
108
Valve Installation
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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Description and Installation
Injection Valve
Object Manager
Prerequisite for the LC Injector HTC/HTS models is Firmware level 4.1.X or
higher, PAL Loader software version 2.1.0 (or higher) and the PAL Object
Manager software version 2.2.1 (or higher) with PAL-xt Object Lists Rev. A.
PAL-xt Object list Parameters:
• Object Name: PAL VlvDrv-5067-4123_1
• Active: +30 °C
• Standby: -30 °C
Definitions:
• Active or Load: Fill sample Loop (Loop in low pressure position)
• Standby or Inject: Load sample from Loop to column (Loop in high pressure
position)
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Description and Installation
Injection Valve
Valve and Actuator Positions
Table 29
Rotor position, view from backside of valve body (actuator side).
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Figure 54
Table 30
Figure 56
110
Valve position “Standby”
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Figure 55
Valve position “Active”
Valve Drive Actuator, View from front of Actuator.
Actuator Position “Standby”
Figure 57
Actuator Position “Active”
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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Description and Installation
Injection Valve
Spare Parts
p/n
Description
5067-4123
Valve (complete) 2 PS/6PT 1200 bar
5068-0029
Stator 2PS/6PT 1200 bar
5068-0030
Rotor Seal, 2PS/6PT 1200 bar
5188-6478
Needle Seals, Rheodyne Valve (10/pk) FEP tubing with SST ferrule, Needle Gauge
22 (OD 0.72 mm)
5068-0031
2 µL Loop
max operation pressure 1200 bar
5068-0032
5 µL Loop
max operation pressure 1000 bar
5068-0051
10 µL Loops
max operating pressure 1000 bar
5068-0033
20 µL Loop
max operating pressure 1000 bar
5068-0034
50 µL Loop
max operating pressure 600 bar
5068-0035
100 µL Loop
max operating pressure 600 bar
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Description and Installation
Injection Valve
Valve Needle Guide
All valves are equipped with a special valve needle guide fitting. This fitting
has a wide diameter to mate with the syringe needle guide on the injection
unit. The valve needle guide also has a countersunk hole to facilitate insertion
of the syringe needle into the injection port.
Figure 58 on page 112 shows the example of the setup with the standard valve,
Cheminert type.
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Figure 58
Valve Needle Guide and Valve Needle Seal
The valve needle guide holds the needle seal. The needle seal, which is a short
length of FEP tubing, forms the seal around the syringe needle. A stainless
steel ferrule is tightened around the FEP sleeve to ensure a leak-proof fit.
112
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Description and Installation
Injection Valve
When
The interval to replace the Needle Seal depends on the number of penetrations. Two factors may be
considered for the decision to replace the Needle seal:
• Leak, seal not reliable
•
Remove the Needle Seal from the inlet port and insert a syringe manually with the corresponding
needle gauge (standard Gauge 22 OD 0.72 mm). Move the needle below the seal point of the
ferrule. If no resistance is observed at this point, replace the Needle seal.
Carry-over
In case of any doubt of possible contamination of the Needle Seal with a compound, replace the
Needle Seal. Contamination can occur if the sample solution is accidentally dispensed too fast
into the valve system. The restriction of valve and loop may cause backpressure and the sample
solution can be forced backwards into the inlet port of the valve.
Parts required
NOTE
#
p/n
Description
1
5188-6478
Needle Seals, Rheodyne Valve (10/pk) FEP tubing with SST ferrule, Needle
Gauge 22 (OD 0.72 mm)
LC Injector HTC/HTS Firmware (level 2.4.X or higher) provides a counter for the number of
injections performed with the LC Injector HTC/HTS System. This counter monitors the
valve switches and not the number of penetrations into the injection port, e.g. for cleaning
the valve an extra penetration is required. It is advisable to monitor this counter in
conjunction with the test for the tightness of the Needle Seal as described above.
• After running routinely a few weeks, the warning limit for the “Inject Counter” gradually
becomes more evident and will be a valuable tool for the user in routine work.
• Path: Menu > Info > Maintenance > Inject Limit (“Inject Counts” reflects the actual
number of injections).
NOTE
To ensure reproducible sample injection and minimize carry-over it is critical that:
• The valve needle guide and the valve needle seal be installed properly,
• The valve needle seal be changed on a regular interval, details see below,
• The needle penetration depth be set accurately.
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Description and Installation
Injection Valve
To specify a limit number of maximum penetrations into the Needle Seal is
not practicable. It is advised to check the tightness of the seal daily or
weekly when first using the LC Injector HTC/HTS System to gain
experience with the specific use in the laboratory related to the
application.
The following simple test provides assurance of tightness:
1 Take the Needle Seal out of the valve port.
2 Manually insert a syringe needle gauge 22 (or Gauge 22S) from top down
into the tube, passing the sealing point of the ferrule.
3 If a restriction is observed, continue to use the Needle seal. If not, replace
the Needle Seal.
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Description and Installation
Interfacing the LC Injector HTC/HTS to other Devices
Interfacing the LC Injector HTC/HTS to other Devices
Synchronization and Output Signals
Synchronization Signals (Sync Signals) are inputs that tell the LC Injector
HTC/HTS when to wait or proceed with a sample-processing step. Output
Signals (Out Signals) are sent from the LC Injector HTC/HTS to external
devices to indicate status or completion of particular processing steps. These
signals are classified as Objects. Physical Events (e.g.TTL-In1) are also Objects
and may be assigned to named signals. LC Injector HTC/HTS Cycles require
that certain signals such as "Start", "Inject", and "Injected" be defined.
See Table 31 on page 116 for the LC Injector HTC/HTS Events and signal
assignments associated with the standard LC Injector HTC/HTS "LC-Inj" Cycle.
Certain types of integration of the LC Injector HTC/HTS system into data
handling or control software, such as ChemStation, Masshunter or EZChrom
accomplish synchronization of the Ready/Start signal directly via software
control (RS232/LAN). A synchronization cable is not required and the Sync
Signal “Start” must be set to “Immediat”.
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To assign a physical Event (e.g. SW-Out 1) to an existing signal Object
(“Injected”) complete the following menu selections:
LC Injector HTC/HTS is shipped with all Cycle events pre-defined as shown in
the table below. If a different physical signal needs to be used, then it will be
necessary to assign a new Event to the Object signal.
If a signal is to be ignored, then its corresponding Event must be set to
“Immediat”.
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Description and Installation
Interfacing the LC Injector HTC/HTS to other Devices
Table 31
Standard Assignment
for LC Injector
HTC/HTS Injection
Cycles
Description
Default Events
Start JobQueue (Sync
Signal)
Start a Job Queue
(Job). Prerequisite is a
defined “Job” with a
method assigned.
Useful for automated,
unattended, timed
sampling.
Immediat
Start (Sync Signal)
Start a cycle (Input
from LC or data
system)
Immediat1
Start2 (Sync Signal)
Continue the cycle
“GC-Dual” for the
second sample
Immediat (Ignore)
Inject (Sync Signal)
Inject READY to
INJECT
Immediat (Ignore)
Inject2 (Sync Signal)
Inject the second
sample for “GC-Dual”
cycle
Injected (Out Signal)
Activated at the
moment the sample
has been injected
SW-Out1
Injectd2 (Out Signal)
Activated at the
moment the second
sample has been
injected
Off
Running (Out Signal)
Active as long as the
Job Queue is being
processed and the LC
Injector HTC/HTS is
not in an error state
Off
1
116
Standard Assignment for LC Injector HTC/HTS Injection Cycles
INTERFACE1 Pin #
7
8(GND)
3
4
If used to synchronize with Sync cable, change setting to “TTL-In1”
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Description and Installation
Interfacing the LC Injector HTC/HTS to other Devices
3
If an item of the class “Sync Signal” is assigned to an output signal, such as
“TTL-In1”, then this TTL contact must be defined as either “Active High” or
“Active Low”, as expected by the HPLC system. This is defined in the Firmware
class “Events”.
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Select the corresponding item and set the mode accordingly.
The setting is provided with the schematic description of the LC Sync Cable if
a dedicated cable is ordered.
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Description and Installation
PAL Software Program
PAL Software Program
PAL Loader Program
The purpose of PAL Loader Software is to create a backup file of the complete
PAL System Firmware. The core software (firmware) and all PAL Firmware
Objects, including all settings (Items), the local methods, and jobs are saved in
one file.
The system requires PAL Loader software, “Loader” version 2.1.X or higher,
and PAL Firmware 4.1.X or higher. Earlier revisions of the loader software
and of the PAL firmware are not compatible. .
NOTE
The new PAL Loader Software is backwards compatible. The software can be used for all
PAL Firmware versions.
The name of the *.exe file has been changed intentionally to avoid conflicts
when old and new versions are installed on the same computer:
• PAL Loader version 1.1.1: PALLOAD.exe
PAL Firmware version up to and including 3.X.X
• PAL Loader version 2.1.X: Setup PALloader (.exe)
PAL Firmware version as of 4.1.X but backwards compatible with lower
versions
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Description and Installation
PAL Software Program
PAL Object Manager Software
The PAL Object Manager software is used to load a new Firmware Object to a
system after a new module (hardware module) has been added to the PAL
System.
The PAL Object Manager software is the software used to handle the so-called
PAL Object List.
The software version 2.2.1 or higher has been developed for the PAL-xt System
that, in addition to serial communication, also permits LAN communication
(TCP/IP). The software is backwards compatible and can be used for all PAL
Firmware versions.
The PAL Object Lists have to match the corresponding PAL Firmware version,
as shown in the list below:
• PAL System Firmware Versions 2.X.X
• PAL Object Lists Rev. K (or higher)
• PAL System Firmware Version 3.0.X
• PAL Object Lists Rev. K (or higher) See specific Object Lists for this FW
Version 3.0.X.
• PAL-xt System Firmware Version 4.1.X (or higher)
• PAL-xt Object Lists Rev. C (or higher)
The two Object Lists for the PAL and PAL-xt Systems can be loaded with the
same PAL Object Manager software. This simplifies the handling in cases
where the PAL and PAL-xt Systems are in use within the same working
environment.
Detailed information is available in the ‘Addendum to PAL User Manual PAL
Object Manager Software’.
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Description and Installation
Establishing Communication with Computer
Establishing Communication with Computer
The System allows a serial communication protocol with a PC or LAN
communication, also referred to as TCP/IP or Ethernet.
The LC Injector HTC requires a dedicated APR Control-xt board and PAL
Firmware level 4.1.X or higher.
COM Port Settings
The port settings are normally defined in:
Windows\Control Panel\Device Manager\Ports.
The standard settings for the serial communication are shown in Figure 59 on
page 121. It is important to understand that the application (e.g. PAL Loader
Software) does actively set the communication parameters, and the baud rate
is set for optimized use.
120
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Description and Installation
Establishing Communication with Computer
Figure 59
COM Port Settings
A quick way to check whether serial communication with the System can be
established is to use the PAL Loader software.
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Description and Installation
Establishing Communication with Computer
Setting up LAN Communication
1 Select the firmware “Communication” class by the following path: Menu/
F3-Setup/Communication
The system automatically searches for the MAC address when the “Setup
Communication” window is accessed.
Figure 60
Reading the MAC Address from the Ethernet Communication Port
This search for and reading of the MAC address takes approximately 10 to
15 seconds. If successful, the MAC Address is stored in the system firmware
and can be viewed on the terminal as follows.
122
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Description and Installation
Establishing Communication with Computer
Figure 61
3
Communication Settings
2 Pressing function key “F1”, “Set Comm” sets the selected communication
mode. The same is achieved by pressing “ESC” or “Home”.
Figure 62
Setting Communication Mode
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Description and Installation
Establishing Communication with Computer
Explanations for the Various Settings for the Communication Protocol
Table 32
124
Explanations Communication Protocol
Parameter
Explanation
Remarks
CommPort
Setup for serial (SER1) or LAN
communication.
If serial communication is selected, all
other parameters in PAL-xt Firmware
“Communication” class are ignored.
DHCP
= Dynamic Host Configuration
Protocol.
If set to “Off”, the IP address is
provided by the user, see IPAddr1 to 4.
The user can ask the network
administrator for a suitable IP
address. If set to “On”, an IP address
is dynamically sought throughout the
provided network.
It is advisable to use a fixed IP
address, set the “DHCP” parameter to
“Off”.
In this mode the PAL-xt System
always has the same fixed IP address.
Troubleshooting is easier for the
network administrator. See also
special remarks below.
IPAddr1 to 4
Enter the IP address divided into four
blocks, example IP default Address:
192.168.0.1.
The IP address is a fixed PAL-xt
System address. The IP address must
be provided by the network
administrator.
SubNet1 to 4
SubNet mask for network
configuration.
The SubNet mask must be provided by
the network administrator.
TCPPort
This parameter is read only, no
changes possible.
The network administrator must be
informed about this setting. The port
must be cleared of firewall protection
to enable communication.
StdGateway 1 to
4
If Gateway is used in network, the
Gateway IP address (switch, hub) has
to be entered.
Gateway is used if independent
networks need to communicate
interactively. Communication within
the network is provided by a hub or
switch, defined as “Gateway”.
MACAddr1 to6
The unique MAC Address for the
PAL-xt System is read and entered
into this PAL-xt communication
protocol.
This is a read only parameter field.
The MAC address is provided by the
Ethernet communication hardware.
The address is also visible beside the
readout from the terminal on the label
affixed to the Ethernet module on the
PCB “APR module”.
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Description and Installation
Establishing Communication with Computer
Special Remarks to the above listed Parameters
DHCP:
In current Firmware level 4.1.X, certain limitations accompany the “DHCP”
communication protocol. If “DHCP” mode is active, a dynamic search for the IP
address is activated. If a free IP address is found, the communication from the
PAL-xt System to the network becomes established. The new IP address
(found through “DHCP” search) is not entered into the PAL-xt Communication
protocol as seen from the local terminal. The parameters “IPAddr1 to 4” are not
updated.
How to overcome this situation is described below with the PAL Loader
software as example but analogously applies the described path to other
application as well.
The Setup for the PAL Loader software requires entering the IP address or
hostname. The dynamically provided IP address cannot be displayed on the
local terminal. To overcome this difficulty, the following setup procedure can
be used:
Preparations
PAL-xt setting in “Communication” class: DHCP = On.
1 Open the “Setup” window and enter the following name in the “Hostname or
IP Address” field: CXXXXXX. The “Xs” must be replaced by the last six
characters of the MAC address, which is known from the PAL-xt System as
parameters “MacAddr4”, “MacAddr5”, and “MacAddr6”.
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Description and Installation
Special Functions
Special Functions
This section describes special functions which can be activated by the “PAL
Extended User Mode”.
The purpose of having two software access levels is to display discrete items
and sections of the firmware at each level. Settings which need to be done
rarely, perhaps at the time of installation of the System, are hidden at level
1(User Level) and revealed at level 2 (Extended User Level). This also protects
the System in group use. Nevertheless, the important points should be known
by the user and are described below.
How to Access “Extended User Mode”
Extended User Mode can be accessed by the following path:
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;("HZije
The above path as shown is an abbreviation for the following steps:
1 Menu/ scroll the cursor to Setup/
2 Press “F3” /
3 Activate “Setup” by pressing the center of the selector knob.
In this User Manual or in technical notes, only the following will be referred to:
Menu > F3-Setup.
126
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Section “F3-Setup”
The path to access this section is shown in Figure 63 on page 127. This “Setup”
section at the Extended User level provides various options to access other
classes which are not visible at the User level. Some classes are known but
may show some new items. Details are described in Table on page 128.
BZcj
;("HZije
HXgdaaYdlc
Figure 63
Selecting “Setup” in Extended User Mode
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Description and Installation
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Object
Description
Ident
The “Identification” of the system provides the opportunity to assign
Users, site, and system names. The serial number of the System can be
entered at this level. Remark: After loading a fresh Firmware Backup file,
the SNo. is displayed as “XXXXXX”.
Access
Access can be selectively opened or limited to different users for the
“Job”, “Method” or “Setup”.
Sounds
The “Beep” sound can be adjusted to specific needs. For safety reasons it
is advisable to keep the default settings, especially the “Warn Move”,
turned ON to signal the start of the System to the user.
Time
The internal clock can be set. Remark: If the clock is fast, reset all items,
from “Year” to “Seconds” and use the mandatory “F1” “Set Time”.
Communication1
The communication mode, serial or LAN, can be selected and conditions
are defined.
System2
Basic parameters can be predefined for specific needs. Details see
“System” on page 129.
Service2
Basic service tests can be activated. Details see “Service” on page 132.
Objects
Accessing the Firmware class “Objects” in the Extended User mode
provides more detailed possibilities for object items. At this level an Object
can be copied or deleted by using the specific function keys. This provides
quick access to add e.g. a second or third injector.
1
Available beginning with PAL Firmware Version 4.1.X.
2
Explained in detail below.
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System
The following “System” Items can be changed by selecting the particular item.
Beginning with Firmware version 4.1.X, an intermediate step was added.
After selecting “System”, the new “Settings” level follows. Activating “Settings”
opens up the described Items below.
Item
Description
Syr Detect
The syringe detection can be turned off in cases of sensor failure or if a
special syringe with unknown ID is installed.
Turning OFF the syringe identification sensor automatically displays a
dialog box enabling the selection of syringe identification manually.
Start Ref
This item provides the choice of referencing all axes at the start of the
system or suppressing the referencing. It is advisable to use this function
actively to detect a possible loss of steps.
Stop XY Error
This function allows turning off the automatic recovery of the system after
detection of a collision (loss of steps of a stepper motor).
Stop XY Error = ON: Automated recovery turned OFF
Stop XY Error = OFF: Automated recovery turned ON
In Firmware version 2.3.X the new feature that recovers automatically after
detecting a collision has been implemented. Firmware version 2.6.2
provides the possibility of disabling automated recovery. If recovery is
turned OFF, the system always checks its position before moving to the
injection port for injection. This can lead to an extra move, crossing over
the X-,Y-axes sensors – if the injection port is at the opposite end of the
X-axis relative to the sample location, (zero X-axis position in-between
sample location and injection port).
For critical application fields, e.g. clinical analyses, it is advisable to turn off
the automated recovery; Stop XY Error= ON.
PlgPathCheck1
The plunger travel path is monitored to ensure that the plunger tip reaches
the syringe zero point in all cases – compared to original plunger
referencing and defining the syringe zero point.
If an error is detected, the System stops operation.
This feature is useful if sample solutions with particles need to be handled,
or if the syringe type is not adequate for the solution composition, or if the
syringe plunger is blocked or even bent, such an error will be recognized
and the system will come to an emergency stop.
PlgChnge Pos
Changing a syringe using the Menu function “Chnge Syr” moves the
plunger up to the position specified by this item. For normal use, accept the
default values.
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Item
Description
Init Syr at
The syringe plunger can be initialized (referenced) at “Home” or “Waste”
position.
If samples with a high danger of toxicity are treated, it is advisable to use
the “Waste” position for referencing.
Inj Signal
An Autosampler is usually the master in a total chromatographic system.
The other components send a “Ready” signal to enable a start of the
Autosampler. At the moment of injection a Start signal is sent from the
System to the other component(-s) of the entire system. With this “Inj
Signal” item the status at which the signal is sent out can be defined. The
syringe is filled with the defined sample volume.
PlgUp = Start signal at the moment the plunger starts moving down for
injection.
PlgDown = Start signal at the moment the plunger reached zero point from
syringe (recommended for large volume injection, but not in combination
with Agilent Chemstation).
ValveSw = Start signal sent at the moment the injection valve is switched,
HPLC technique.
PreInj = Start signal sent at a time before injection, as with a sample
preparation device started before a GC or LC system. Minus time relative to
chromatographic start time.
Vial PrePress
This item can be activated to avoid a vacuum if a relatively large amount of
sample is taken out of a small volume vial. If e.g. a volume of 50 µL is
specified for injection, the PAL system first takes 50 µL ambient air and
“injects” this volume into the sealed vial, causing overpressure in the vial.
After this step, the sample volume can be aspirated without forming a
vacuum.
This functionality is only valid for liquid sampling, not for the Headspace
technique.
Starting with Firmware version 4.1.X, this item is also active in combination
with Cycle Composer.
LC-Inj
Standard cycle for HPLC technique.
Details on method parameters see “Injection Cycles” on page 196” and
“Software Flow Chart” on page 246.
LC-Cut
Cycle for column switching with two valves, HPLC technique.
Both valve drives have to be controlled through “AUX” interfaces.
Details on method parameters see “Software Flow Chart” on page 246.
GC-Inj
Standard Cycle for GC technique.
GC-InS
Standard cycle covering GC Sandwich technique.
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Item
Description
GC-Dual
Cycle for GC technique which allows injecting two samples in two different
injectors. The start signal is sent after injection of both sample solutions.
HS-Inj
Standard cyle for GC headspace technique.
SPME
Standard cycle for GC SPME technique, Solid Phase Micro Extraction, Fiber
technique.
TTS Sotax
The two cycles are dedicated cycles for “dissolution applications”. TTS
cycle is used for transdermal patches and the Sotax cycle is used for tablet
dissolution.
1
Available beginning with Firmware version 4.1.X.
NOTE
Standard injection cycles are selectable and activated directly from the System. Any
deviation from a standard cycle, a Cycle Composer Macro, or Cycle Editor (ICC-CE) Cycle
must be programmed for customized requirements.
NOTE
In Firmware versions < 4.0 the standard cycles are grouped in Item “Cycles”.
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Service
“Service” items may be changed by selecting the particular item (see Table on
page 132).
Item
Description
Check Motors
Every stepper motor can be moved separately. This function is useful to
check out a specific motor or to perform an endurance test after a repair.
The Start and Stop positions of the test path can be selected. The motor
Current, Speed, and Acceleration are selectable (accept default value to be
sure). A Pause time can be specified. A Cycle Limit and Actual Cycle
Counter are useful tools for endurance tests.
Phase A: This item is used to verify the functionality of the stepper motor.
The current can be manually increased for tests. Used for troubleshooting
only.
Phase B: This item is used to verify the functionality of the stepper motor.
The current can be manually increased for tests. Used for troubleshooting
only.
MicroStep Inx: The motor can be moved within a range of up to four full
steps by applying the assigned current.
NOTE
These items are typically used for service. If applied, do not leave
the test settings for longer than a few minutes. If current setting is
in a critical range the motor can be damaged.
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Item
Description
Test Head
This tests the functionality of the Injection Unit (“Head”). Two tests can be
activated by the function keys:
“F1” Check Plg: The stepper motor of the plunger drive is tested. Sensor
check.
“F2” Check Z: The vial detection sensor and “Needle Guide Blocking”
function is tested.
These tests are only required if any erratic behavior of the System is
observed. In such cases contact a representative of Agilent Technologies.
Test Switches
X-Limit = Actual status of X-axis sensor. Move the X-Y-Carriage manually
across the limit switch to verify activation of the sensor.
Y-Limit = Actual status of X-axis sensor. Move the Y-axis manually to the
Y-zero point (at X-axis), crossing over the limit switch to verify activation of
the sensor.
Z-Limit = Actual status of Z-axis sensor. Move the syringe slider assembly
manually up to the zero Z-axis point, crossing over the limit switch to verify
activation of the sensor.
Aux1 Limit = Actual status of Aux1 limit sensor
Aux2 Limit = Actual status of Aux2 limit sensor
Plg –Pos = Actual status of Plg –Pos
Needle Guide = Actual status of needle guide sensor
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Section “F3-Setup” / Objects
The same as is described above applies for the section “Objects” opened in
Extended User Mode. New classes are visualized and known classes may
contain new items accessible only on the level of the Extended User Mode.
To open this level, use the following path:
BZcj
;("HZije
DW_ZXih
HXgdaaYdlc
Figure 64
134
Selecting “Objects” in Extended User Mode
Object
Description
Syringes
At this level all loaded syringes are selectable. At the “Utilities” user level only
the active syringe is visible.
Vials
The Firmware Objects accessed on the Extended User Mode have more specific
items selectable than at the User level. Tolerances or ranges such as, e.g.
“Maximum Penetration” can be defined and limited for the user such that this
value cannot be exceeded. In the Object “Tray Types”, details for the various
Items are described more closely.
Tray Types
The object class “Tray Type” is only accessible via the Extended User Level. All
loaded Tray Types are visible and selectable. Details, see “Tray Type” on
page 138.
Tray Holders
The Firmware Objects accessed on the Extended User Level have more specific
items selectable than at the User level. Depending on the type of Tray Holder,
e.g. an Access and Restore Path or a Heater and Agitator (motor) is assigned.
Typical example, the Tray Holder “Agitator”.
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Object
Description
Trays
All loaded Trays are visible and accessible. A Tray must be assigned to a Tray
Holder. A “TrayTypeGroup” identification checks if the specified Tray Type is
allowed in this combination. The offsets X-,Y-, Z and the inclination corrections
for a row or column are explained in the “Utilities Functions” section.
The “Path Offset X-,Y-, Z” is used only if a “Path” is assigned to the specified
Tray Holder, typically a Stack or Agitator uses a Path.
Wash Stations
The Wash Station and its positions “Wash1” and “Wash2” are visible and
selectable. Details , see “Wash Stations” on page 144.
Injectors
The Firmware Objects accessed on the Extended User Mode have more specific
items selectable than at the User Level. Tolerances or ranges such as
“Maximum Penetration” can be defined and limited for the user so that this
value cannot be exceeded.
In the “Tray Types” Object, details for the various Items are described.
Furthermore, in this Firmware Object class a “Valve” and/or a “Heater” can be
assigned.
Note that the Wash Station Positions “Waste” and “Waste2” are listed in this
“Injectors” class.
Valves
The Valves and Valve Drives connected and controlled by an “AUX” interface
are listed in this Object class. For more details, see the Valves and Valve Drives
Addendum.
Note that the valves controlled by serial communication (Serial Valves or
Multiposition Valves) are in the Firmware Object class “Serial Valves”.
Dilutors
The Firmware Objects accessed in the Extended User Mode have more specific
items selectable than at the User level. Tolerances or ranges such as
“Maximum Fill Speed” can be defined and limited for the user so that this value
cannot be exceeded.
Items such as “Prime Volume”, “Waste to”, “Motor Drives”, or “Scale Length”
are specific items that remain hidden at the User level.
Positions
The various positions used with the System are selectable at this level.
Positions such as “Home”, “Change Syringe”, or a Path Point, reference the
Paths for a specified Tray Holder directly, such as “Agitator” or “Stack”.
Selecting an item, e.g. “Home” will provide “F1”, “Check Pos” on the screen.
Activating this function provides the possibility of verifying X-, Y-, Z-axes values
for this particular position.
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Object
Description
Paths
A Path defines specific movements for the Injection Unit, which represent a
repetitive task assigned to a Tray Holder. Typical examples are opening and
closing a drawer from a Stack or opening the lid of an Agitator.
The Path is composed of several steps (points) which are added up in a
sequence. Each individual step (point) can be positioned by teaching X-, Y-,
Z-axes. In most cases the return path is in the reverse order of the starting path.
Example “AgiOpnL” and “AgiClsL”. If one setting of a point is changed, this
change will be valid for the return path as well.
Sync Signals
The various synchronization signals are described in “Synchronization and
Output Signals” on page 115. At this level no other functionality, as shown at
the user level, is provided.
Out Signals
The various Out-Signals are described in “Synchronization and Output
Signals” on page 115. At this level no other functionality, as shown at the user
level, is provided.
Events
The various synchronization signals are described in “Synchronization and
Output Signals” on page 115. At this level no other functionality, as shown at
the user level, is provided. Three events are newly available at this extended
user level:
Pwr-Out1, Pwr-Out2 and FlushVlv. The main purpose of these three power-out
signals is to activate a solenoid, such as for the Wash Station or the Gas Flush
Valve. At this level the functionality of the signal can be tested.
Serial Valves
Serial valves, or “Multiposition Valve Drives” are powered and controlled by the
serial (RS232) control and not through the “AUX” interface. With firmware level
4.1.X a “Mode” is now selectable. The mode refers to the setting of the Valve
Drive, whether the current for the motor is set to “High Speed” or “High
Torque”.
For more details, see Addendum to PAL User Manual “Valve and Valve Drive”.
Out Exp Box
The “Out Expansion Box” provides 8 TTL contacts and 8 relay contacts (24 V
contact closure). The optional module (box) is connected and controlled by
“Interface 2”.
For more details, see Out Expansion Box Addendum to PAL User Manual.
RS232
This Object allows selecting the serial ports for the specific needs. Choices are:
Remote, Terminal, Printer, Barcode, ExtDev, VICI-Vlv (for serial valve drives),
or None.
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Object
Description
Motors1
In the class “Motors” the Objects “Motor-Y” and “Motor-Z” are accessible to
enable the user to select specific motor parameters in case a specific
application requires an adjustment.
Motor-Y Strategy Three different settings are selectable for the Y-Motor
movement strategy:
• Rectang = normal movement as used for system. (In firmware version <
4.1.X. this mode was called “Auto”.)
• XY-Simul = X,-Y-axes move simultaneously, which allows movement in a
diagonal direction. (At firmware version < 4.1.X this mode was called
“Rectang”, but it was not active at lower versions.)
• Retract = Y-axis moves from any point first to zero Y (back to the X-axis)
before the X-movement is started. This mode allows bypassing an object
(such as a large GC detector) which may be in the travel path if the default
mode “Rectang” is used.
• Path Speed = Y-axis motor speed in conjunction with a “Path”, example:
Opening and closing a drawer of a Stack. In certain situations it is necessary
to slow down the speed to open and close a drawer. Typical application
example is fraction collection, collecting directly into a well plate. If the
liquid level is high, the liquid can wash over into the next row if drawer
movement is too fast.
NOTE
At Firmware level 4.1.X these items of the Y motor have been made
available at the Extended User Level.
Motor-Z For the ‘Motor-Z’, following speeds are selectable:
• Penetr Speed = This is the speed used to penetrate the syringe needle into
any object, with the exception of the injector. Example: Vial penetration.
• Inj Penet Spd = Injector Penetration Speed. With PAL Firmware version 2.X
and 3.0 the injector penetration speed is identical to the injector retraction
speed.
• Inj Retr Spd = Injector Retract Speed. With PAL Firmware version 4.1.X, the
injector speed was separated into a penetration and a retraction speed. This
allows safe penetration of a vial by any type and diameter of syringe needle
without bending through the septum of the GC injector. The needle
retraction speed can be increased to speed up the entire injection process.
This new feature enables the user to avoid uncontrolled evaporation of any
remaining sample solution in the needle.
The visibility of the Motor-Z parameters at the Extended User Level was
introduced with PAL Firmware version 4.1.3.
1
Available beginning at Firmware Level 4.1.X
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Tray Type
The following object “Tray Type” Items may be changed by selecting the
particular item. First select the desired Tray Type, such as “VT32-20”.
NOTE
The definition of Row and Column is given in “Definition of a Tray Row and Column” on
page 94.
By reversing the first six entries, the order of injection can be changed.
Example: Tray Type “VT32-20” has a “Row Length Y” and a “Col Length X” defined.
Reversing to “Row Length X” and “Col Length Y” would start with sampling from left to
right, as used e.g. with Tray Type “DW96”.
The values of the items “Spl Per Row” and “Spl Per Col” must also be reversed.
138
Item
Description
Row Length X
Defines the Row length in the X-direction from the center of the first to the
center of the last vial position, measured in mm.
Row Length Y
Defines the Row length in the Y-direction from the center of the first to the
center of the last vial position, measured in mm.
Col Length X
Defines the Column length in the X-direction from the center of the first to
the center of the last vial position, measured in mm.
Col Length Y
Defines the Column length in the Y-direction from the center of the first to
the center of the last vial position, measured in mm.
Spl Per Row
Number of samples per Row.
Spl Per Col
Number of samples per Column
Pattern1
The pattern of the Row or Column arrangement has to be defined.
• Regular = square arrangement (standard)
• Staggrd+ = Rows are arranged in staggered pattern.
• Offset of second Row is shifted by +50 %5 of hole pattern.
• Staggrd- = Rows are arranged in staggered pattern.
• Offset of second Row is shifted by -50 % of hole pattern.
See Graphics below.
TrayTypeGroup
Software protection so that not every Tray Type can be placed on any Tray
Holder. Example: VT32-20 cannot be fit into a Stack.
Plate Thickn
Thickness of Tray plate bottom (not the total height of the Tray). Measured
in mm.
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Item
Description
Vial Height
Total height of the vial including cap to seal if applicable. Measured in mm.
NOTE
The sum of the plate thickness and vial height adds up to the total
height This is the point where the Injection Unit expects an object.
Vial Trnsprt2
The mode to transport a vial is defined by:
• None = No transport required. Example liquid sampling.
• Magnet = Magnetic Transport. Requires magnetic vial caps. The
Injection Unit moves to the side to slide the vial off when moved away.
• Needle = Device is transported with the syringe needle inserted.
Example: special vials where a magnetic cap cannot be crimped, e.g.
Vacutainer.
ZSlideOffRetr1
If magnetic transport is selected, the distance can be defined to move the
syringe slider up (Z-direction), lifting up the vial by magnetic force.
Measured in mm.
YSlideOffDist1
If magnetic transport is selected, the distance can be defined to move the
Injection Unit to the side, Y-direction, to slide the vial off. Measured in mm.
Barcode
If a Barcode Reader is installed, the mode has to be set for AutoFix.
None = No Barcode Reader option requested for this Tray Type.
Z Tolerance
A tolerance window to give a plus/minus range (expressed in mm) where
the Injection Unit must expect an object.
If the value of this item is > 0, this is a relative detection mode and the
sensor from the Injection Unit needle guide is active.
If the value of this item is set to 0 the sensor of the needle guide is turned
off and the syringe slider moves to an absolute value, Z-axis position as
specified in the object.
Z Retract
The distance to lift up the syringe slider before the Y-, Z-axes assembly
moves across the unit can be specified, expressed in mm.
Max Penetr
The maximum allowed Needle Penetration can be defined as a safety so
that the user cannot destroy the needle tip by going too far down. The Tray
Type is prepared for a specific vial size; this value is therefore unique to this
combination. The maximum needle penetration must not exceed the length
of the syringe needle, considering the loss due to needle guide, etc.. Typical
needle length is 51 mm, maximum penetration must not exceed 48 mm.
Needle Penetr
This item is identical to that used on the User Level or, if available, as a
method parameter of a local cycle. The value defined on these levels is
mirrored to the Extended User level.
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NOTE
Item
Description
Spl1 Offset X
The X offset is defined as the distance from the outer edge of the Tray in
the X-direction to the center of the vial position 1. Measured in mm.
Spl1 Offset Y
The Y offset is defined as the distance from the outer edge of the Tray in
Y-direction to the center of the vial position 1. Measured in mm.
Spl1 Offset Z
The Z offset can be selected for special cases where, e.g. the syringe
needle has to reach a deeper point before penetration into a vial. A Z offset
value = 0 considers the Tray surface or vial top as the reference point.
Measured in mm.
1
Available beginning with Firmware version 4.1.X
2
Available beginning with Firmware Level 2.5.X
The dimensions used for Row/Column Length or X-, Y-, Z-Offsets for Sample Number 1, are
specific for a particular Tray Type. If a general adjustment for a Tray from a specific vendor –
e.g. for a Deepwell Plate – has to be made, this should be done in the “Tray Type” class of
Object. For fine tuning, the Offset items from the class “Trays” can be used.
Explanation for various Patterns in Tray Type:
1 Pattern Type “Regular”.
The sample or well positions are equal distributed.
Gdl
8dajbc
8dajbc
Gdl
Figure 65
Pattern Tray Type “Regular”
2 Pattern Type “Staggrd+”.
The sample or well positions are arranged in a staggered pattern. Offset of
second Row is shifted by +50 % of hole pattern.
140
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3
Gdl
8dajbc
8dajbc
Gdl
Figure 66
Pattern Tray Type “Staggered+”
3 Pattern Type “Staggrd-”.
The sample or well positions are arranged in a staggered pattern. Offset of
second Row is shifted by -50 % of hole pattern.
Gdl
8dajbc
8dajbc
Gdl
Figure 67
Pattern Tray Type “Staggered-“
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Trays
The following object “Tray” Items can be changed by selecting the particular
item. First select the desired Tray, such as “Tray1”.
Item
Description
Tray Holder
The Tray Holder to which the Tray is assigned is specified.
TrayTypeGroup
All Tray Type Groups which are allowed for this particular combination of
Tray and Tray Holder are displayed. The TrayTypeGroupfor the Tray Type
itself is specified for each Tray Type.
Tray Type
The Tray Type to be used is linked to the Tray.
This item is usually set at User level in Utilities > Trays. See above.
Offset X
The X offset is defined as the distance from the center of the teaching
position from the Tray Holder to the edge of the Tray in X-direction.
Measured in mm.
This item is available at User level in section Utilities > Trayand can be
used for fine tuning a Tray installed in a Tray Holder. Important for
well-plates 384 installed in a Stack.
NOTE
Position#1 is the only correction point.
Offset Y
The Y offset is defined as the distance from the center of the teaching
position from the Tray Holder to the edge of the Tray in Y-direction.
Measured in mm.
This item is available at User level in section Utilities/Tray and can be used
for fine tuning a Tray installed in a Tray Holder. Important for well-plates 384
installed in a Stack.
NOTE
Position#1 is the only correction point.
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Item
Description
Offset Z
The Z offset is defined as the distance from the plate surface of the Tray
Holder to the teaching point, black needle guide flush underneath plate of
Tray Holder (which corresponds to plate thickness of Tray Holder).
Measured in mm.
This item is available at User level in section Utilities/Tray and can be used
for fine tuning a Tray installed in a Tray Holder. Important for well-plates 384
installed in a Stack.
NOTE
Position#1 is the only correction point.
Path Offset X
The Path Offset X can be specific for a Tray, adjustment in X-direction.
Expressed in mm.
NOTE
This item is only valid if a Path is assigned to the Tray Holder as
specified in this combination.
Path Offset Y
The Path Offset Y can be specific for a Tray, adjustment in Y-direction.
Expressed in mm.
NOTE
This item is only valid if a Path is assigned to the Tray Holder as
specified in this combination.
Path Offset Z
The Path Offset Z can be specific for a Tray, adjustment in Z-direction.
Expressed in mm.
NOTE
This item is only valid if a Path is assigned to the Tray Holder as
specified in this combination.
dxRow1
Correction for inclination of a Tray in X-row direction. For details, see
“Correction for Sample Position” on page 95.
dYRow1
Correction for inclination of a Tray in Y-row direction. For details, see
“Correction for Sample Position” on page 95.
dzRow1
Correction for inclination of a Tray in Z-row direction. For details, see
“Correction for Sample Position” on page 95.
dxCol1
Correction for inclination of a Tray in X-column direction. For details, see
“Correction for Sample Position” on page 95.
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Item
Description
dyCol1
Correction for inclination of a Tray in Y-column direction. For details, see
“Correction for Sample Position” on page 95.
dzCol1
Correction for inclination of a Tray in Z-column direction. For details, see
“Correction for Sample Position” on page 95.
1
Available beginning with Firmware Version 4.1.X
Wash Stations
The following object “Wash Station” items may be changed by selecting the
particular item. First select the desired Wash Station, such as “Wash1”.
144
Item
Description
Position X
X-axis position for Wash Station.
Position Y
Y-axis position for Wash Station.
Position Z
Z-axis position for Wash Station.
Z Tolerance
A tolerance window for a plus/minus range (expressed in mm) where the
Injection Unit has to expect an object.
• If the value of this item is > 0, this is a relative detection mode and the
sensor from the Injection Unit needle guide is active.
• If the value of this item is set to “0” the sensor of the needle guide is turned
off and the syringe slider moves to an absolute value, Z-axis position as
specified in the object.
Z Retract
The distance to lift up the syringe slider before the Y-, Z-axes assembly moves
across the unit can be specified, expressed in mm.
Max Penetrat
The maximum allowed Needle Penetration can be defined as a safety so that
the user cannot destroy the needle tip by going too far down. The maximum
needle penetration must not exceed the length of the syringe needle,
considering the loss due to needle guide, etc..
Typical needle length is 51 mm, maximum penetration must not exceed 48 mm.
Needle Penetr
This item is identical to that at the User level. The value defined at this level is
mirrored to the Extended User Level.
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Description and Installation
Special Functions
Item
Description
Type
The specific mode for the Wash station in use has to be defined:
• Standard = Standard Wash Station. Syringe is filled with wash solvent and
expelled into Waste.
• Pulse = A pulsed signal is sent to activate the device. Used for the “Fast
Wash Station” and “Active Wash Station”. See Type `Active´ for details.
• Flow = Continuous flow of liquid by gravity. No electrical contact required
for a solenoid.
• DLW = Dynamic Load & Wash. Item used to activate specific solenoid for
DLW Wash Station.
• Active = Used for Active Wash Station in conjunction with the Wash
Station Print with PNo. ‘APR WASHPUMP B.A.’ (Revision B). Simplified
print, activation of pumps is controlled by PAL Firmware. Requires Object
Lists ‘PAL WashActive_Rev. B’.
Flow Control
The power source to activate a Wash Station is specified. For the Fast, Active
Wash Station and DLW the Event “Power-Out1” and “Power-Out2” are used.
Ndl Cln Path
A Path can be assigned to the Wash Station.
Waste to
The Waste position is assigned to the Wash Station.
Clean Volume
A percentage of the syringe maximum volume is defined for the cleaning step.
Clean Count
The counter monitors the number of wash cycles. The setting in the method
has the same functionality and the value is mirrored to the Extended User
Level.
Max Fill Spd
The fill speed for the syringe is defined with this item. If the maximum value of
the Fill Speed (10.0 ml/s) is selected, the system takes the injection syringe Fill
Speed as defined in the method.
Max Eject Spd
The Eject Speed is the speed used to eject the wash solvent during the wash
cycle. This speed is usually higher than the Fill Speed. If the maximum value of
the Eject Speed (10.0 ml/s) is selected, the system takes the injection syringe
Eject Speed as defined in the object class “Syringe”.
Rinse Time
Rinse is the time to rinse the wash port with wash solvent after the wash cycle
of the syringe has been finished.
In the case of a Fast/Active Wash Station, the syringe needle is pulled out of
the wash port. The wash solvent flows without restriction. As a result, higher
solvent consumption has to be taken into consideration.
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Troubleshooting
Troubleshooting
NOTE
The autosampler represents always one of several components in a chromatography
system.
The following Troubleshooting Guide is limited to the autosampler only.
No or very low detector signal is observed.
Probable cause
Suggested actions
1 Clogged syringe.
Remove syringe and aspirate/dispense liquid
manually. Clean syringe.
2 Bent needle.
•
Check X-,Y-, Z-Axes positions.
•
Check vial or well-plate septum.
3 No sample liquid is injected.
Check and/or adjust Needle penetration into
sample vial (see “Utility Functions” on page 27).
4 Sample volume too low.
Increase sample volume.
5 The valve needle guide and / or needle seal
Check valve needle guide and seal (see “Valve
Needle Guide” on page 112).
are not properly installed.
6 The valve ports are not plumbed correctly to
the pump and/or detection system.
146
Check plumbing connections (see “Injection
Valve Flow Path” on page 106).
7 Wrong valve type specified.
Check valve type by selecting the path: Menu >
setup > Objects > Injectors > LC Vlv1 > valve.
8 Rotor orientation
Check the rotor orientation. (This possibility is
valid for Rheodyne valves only.)
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Troubleshooting
Mobile phase and/or sample flows out of the wrong ports on LC
injection valve.
Probable cause
Suggested actions
1 The valve rotor is reversed (180 ° out of
Remove the rotor and reinstall in the correct
position. See “Injection Valve / Valve Rotor” on
page 173).
alignment).
Sample is backing up on the valve needle guide.
Probable cause
Suggested actions
1 High restriction in valve flow path.
Check the restriction of sample loop, valve
waste tube or clogged connection (ferrules or
tubing distorted).
2 Valve needle seal leaks.
•
Check needle seal type.
•
Change valve needle seal (see “Valve Needle
Guide” on page 112).
3 The needle penetration depth for the
Adjust the Injection Valve Needle Penetration .
injection valve is not set correctly.
4 The syringe Plunger speed is too high
Reduce Inject Speed in method.
resulting in excessive pressure in inlet.
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Syringe does not fill properly.
Probable cause
Suggested actions
1 DLW Actuator/Solenoid not functioning.
PAL Firmware level < 4.0 or PAL Firmware
Object for ‘DLW Option’ not loaded onto the
system.
2 Solvent path blocked.
Prime the system or inspect it to identify the
reason for clogging.
3 Air bubbles trapped in the system, caused by Ensure tight tubing connections at all points;
loose connections or tubing not cut square.
verify that all tube ends are cut properly.
4 Syringe plunger tip worn out.
Replace syringe plunger. Check if glass barrel is
scratched (damaged). If in doubt, replace the
entire syringe at once.
5 DLW-2 Actuator Control PCB defect.
If possible, verify the DLW Actuator Holder Assy.
on another System. If there is a definite defect,
replace the PCB.
6 DLW-2 Holding Loop.
Check the DLW-2 Holding Loop. Is the solvent
flowing freely in the loop?
7 Injection Needle (Holding Loop).
Check the front section of the holding loop. - Is
the needle entry free of debris, not clogged?
Sample peaks or responses are not reproducible.
148
Probable cause
Suggested actions
1 Dirty syringe.
Use the ‘Priming’ utilities as described in
Section ‘Operation’.
2 Syringe pressure differences.
Increase Pullup Delay value.
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Description and Installation
Troubleshooting
Probable cause
Suggested actions
3 Vacuum created in sample vial.
Reduce sample volume in sample vial. Use
setting under F3-Setup > System >
PrePressureVial.
4 Method Parameters
Use the macro default settings and observe the
following crucial parameters first: Fill speed,
Pullup delay, Injection Speed, Post Inj Delay.
Do NOT use the standard injection cycle ‘LC-Inj’.
Please note that this cycle cannot be used for
the DLW operation. However the values of these
parameters can be used as a good starting point.
Peak distortion or tailing.
Peak distortion or tailing
Probable cause
Suggested actions
1 Method Parameters.
See recommended actions above.
2 Solvent for analytical solution not suitable.
Change solvent composition to a lower polarity
(in case of reversed phase chromatography).
3 Tubing internal diameter.
Check tubing internal diameters for connection
tubings and injection loop. See
recommendations as given with poster
‘Tips&Hints for HPLC Technique’ (CTC
Anayltics).
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Probable cause
Suggested actions
4 Bad connections.
Connection tubing’s not square cut or wrong
type of nuts or ferrules installed. See
recommendations as given with poster
‘Tips&Hints for HPLC Technique’ (CTC
Analytics).
5 HPLC related problems.
Any parameter like flow rate, composition of
mobile phase or wash solvent, column selection
(diameter, pore size, etc.) can contribute to BAD
PEAK SHAPE. Check troubleshooting guide from
the HPLC manufacturer or any other source.
Excessive carryover between samples
Probable cause
Suggested actions
1 Loose, unstable or wrong connection.
Check all connections within the DLW-2 System,
such as Holding Loop, injection loop
connections, or valve plumbing connections etc.
2 Inappropriate wash solvent(s).
Use appropriate wash solvent(s).
3 Waste tubing ID at injection valve is too
Replace the waste tubing with larger ID tubing.
small, causing waste liquid to be pulled
back, by capillary action, into rotor groove.
4 Air gaps not formed. Liquids of different type • Verify the DLW method parameter for ‘Airgap
Volume’.
diffusing into each other.
•
5 ID of solvent tube from injection valve to
DLW Wash station is too small.
6 Dirty needle, Holding Loop and/or valve
injection port.
7 Damaged or grooved valve rotor.
150
Prime the entire system.
ID of 0.8 mm (1/32”) is required as a minimum
internal diameter to avoid any restriction.
Use the 'Priming' utilities as described in
Section Operation
Replace valve rotor, see “Injection Valve / Valve
Rotor” on page 173.
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Troubleshooting
Probable cause
Suggested actions
8 Valve needle seal leaks.
Change valve needle seal, see “Valve Needle
Guide” on page 112).
9 Inappropriate wash solvent composition.
Use appropriate wash solvent. Observe also the
order of use for biological samples. The first
wash is always an aqueous solution followed by
organic solvents.
10 DLW Actuator/Solenoid questionable,
Verify the functionality of the DLW Actuator. Is
solvent flowing when active? Use command
“Utilities/Wash Station” for tests.
defect.
11 Solvent Frits in wash solvent reservoir
•
Clean the frits in a ultrasonic bath with an
appropriate solvent.
•
If flow is not ensured after cleaning, replace
frits.
•
Replace wash solvent and clean reservoir
bottle and tubing carefully.
blocked.
12 Inserts of Wash station contaminated or
clogged.
Clean all parts of the DLW Wash Station by
using an appropriate solvent (according to the
application) and if possible an ultrasonic bath.
Unexplained collisions of the Injection Unit.
Probable cause
Suggested actions
1 Object(s) not defined correctly.
Define Object(s) correctly. (See “Defining Object
Reference Positions” on page 88).
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Troubleshooting
“Object Not Detected” (Object e.g. Tray1, LCVlv1, Stk1- 01)
Probable cause
Suggested actions
1 Incorrect Z value for the affected Object (e.g. Redefine Z value for the affected Object. (See
“Defining Object Reference Positions” on
page 88).
LCVlv1, THldr1 or CStk1-01).
2 Missing sample vial, Deepwell- or
Microtiterplate.
Check position and insert, if necessary, vial,
deepwell- or microtiter plate.
“Object Collision before Z Tolerance” (“Object” e.g. Tray1, LCVlv1,
Stk1-01)
Probable cause
Suggested actions
1 Incorrect Z value for the affected Object.
Redefine Z value for the affected object. (See
“Defining Object Reference Positions” on
page 88).
“Invalid Motor Status Motor AUX1”. (AUX2)
152
Probable cause
Suggested actions
1 Injection valve not connected.
Switch OFF LC Injector HTC/HTS and check
connection between LC Injector HTC/HTS and
Valve Drive.
2 Defective LCVlv1 Connection cable.
Change connection cable.
3 Control Board defective.
Replace Control-HTC or Control-xt Board.
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Description and Installation
Troubleshooting
3
Special Considerations for Troubleshooting
Tubing Connections
Special attention must be paid to all tubing connections. Common mistakes in
routine practice are:
• Nut and ferrule not connected tightly and leak
• Pilot distance incorrect
• Tubes not cut square
• Wrong type of ferrule or nut used
• Air bubbles trapped
;ZggjaZ
BVaZcji^ciZgcVa
IjW^c\
E^adi
Figure 68
Definition of Terms for Tubing Connections
Disregarding these basic rules described below could lead to:
• Dead volume
• Peak deformation or peak splitting
• Carry-over effects
• Decreased resolution of the tubing between column and detector
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Troubleshooting
Nut and Ferrule Tightening
Stepwise Tightening
CAUTION
Overtightening
Overtightening even once will damage the seat and the next connection can only be
sealed by force. If the seat is damaged, there is a considerable chance that the ferrule
will stick and cannot be removed without force or special tools (such as a drill).
If different materials are used, such as polymer tubing and nut together with a
stainless steel ferrule, then over-tightening is always a danger. The stainless steel
ferrule will bite into the polymer tube and block the flow.
➔ Do not over-tighten the nut/ferrule. If a connection proves to be leaky, use another
quarter turn to tighten. Step-by-step tightening is the correct approach.
The first important rule when tightening a nut and ferrule is to proceed
stepwise:
1 Tighten the nut by hand as much as possible.
2 Continue to tighten using a wrench until resistance is felt. Add one or two
quarter turns to reach the sealing point.
3 Open the connection and remove the assembly to inspect the pilot distance,
see Figure 68 on page 153.
4 Install the assembly again and tighten the nut manually at first. Use the
wrench to reach the same sealing or resistance point as in the step before
and add one extra quarter turn to obtain the final seal.
5 Check the seal when liquid is pumped through the system. In case a leak is
observed, tighten once more by another quarter turn. The rule is to always
go from leaking to properly sealed, and never try to release an
over-tightened seal.
NOTE
When installing a tube with a narrow bend, typically a loop, it is advisable to fix one
connection first without inserting the second nut into the female counterpart. After
tightening, open the connection. Start preparing the second connection without inserting
the first nut. Open the second connection again and check the pilot distance. The last step
is to insert the complete tubing with the two connections and make a final seal as
described above.
This stepwise installation of a bended tube is mandatory for PEEKsil tubing (fused silica
tubing sheathed with PEEK polymer).
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Re-use of Installed Nuts/Ferrules
NOTE
Do not re-use an installed nut/ferrule for any other connection. Any tube with an installed
nut/ferrule used for one purpose should not be installed for any other application.
1 Cut a portion of the tube end until straight tubing is reached and remove
the ferrule.
NOTE
This rule also applies to loops. Never re-use a loop with another valve or a valve after
exchanging the stator.
NOTE
If a finger-tight nut has been installed, replace it as well.
If a stainless steel nut has been used, ensure that the correct type is used, see “Wrong
Nuts or Ferrules used” on page 159.
NOTE
Newly installed finger-tight ferrules can hold a backpressure of up to 200 bar.
This remark is valid for the well known ‘finger tight nuts’ which have been used for a long
time. Recent developments try to overcome this deficiency. Please observe the latest
developments on the market to find a suitable solution.
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Troubleshooting
Pilot Distance Incorrect
NOTE
When making a new tube connection it is critical to follow these guidelines.
NOTE
Failure to follow this procedure will lead to an incorrect pilot distance, which will result in
actual dead volume (not delay volume).
1 Slip the nut and ferrule over the tube. The end of the tube should stick out
approximately 50 mm (2 inch).
2 Press the tube end firmly into its female counterpart for the connection.
3 Move the nut and ferrule together into the female counterpart for
connection. At this stage is it important to maintain pressure on the tube to
avoid the tube slipping backwards, out of position.
4 Tighten the nut as described above.
Figure 69
156
Pressing the Tube into Place
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Troubleshooting
Tube Not Cut Square
All tubes must be cut absolutely square. Any deviation from square causes a
dead volume which yields carry-over and other chromatographic effects. This
applies to stainless steel, PEEK or Polymer tubing. It is important to use the
correct tool for each material when cutting a tube.
• Stainless steel tubes are often cut using pliers. Often, an egg-shaped profile
will result, which will no longer seal and will cause dead volume. Dedicated
pliers for HPLC tubing are available; nevertheless, it is recommended to use
precut tubing which is cut smooth, clean and passivated.
• Polymer tubes, such as PEEK, PTFE, PFA, etc., are soft and entice the user
to cut them with any handy tool. Commercially available tubing cutters
from many manufacturers are available. If the blade does not provide a
clean, right-angle cut, use a different technique or tool.
The best and most commonly-used method for cutting tubing of any
material is to use a cutter with an adjustable blade.
1 Carry out the initial turns.
2 Readjust the blade a very little.
3 Turn once more.
4 Adjust again until approximately half of the tubing wall is cut.
5 Hold the tube on both sides of the cut with flat-nose pliers.
6 Twist the tube until it breaks.
NOTE
For polymer tubing, the same can be done by replacing the flat-nose pliers with tweezers
with a flat tip.
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Troubleshooting
IjWZZcYXjii^c\
.%•
8dggZXi
6c\aZY
Lgdc\
Gdj\]dgegZhhZYZY\Zh
Lgdc\
Figure 70
158
Illustration of various Tubing Cuts
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Description and Installation
Troubleshooting
3
Wrong Nuts or Ferrules used
Do not use a nut or ferrule from a vendor other than specified for the product.
Figure 71 on page 159 illustrates the resulting dead volume that occurs when
an incorrect type is used for connecting.
Figure 72 on page 159 shows the various forms of the ferrules from different
manufacturers. Be aware that some of them are very similar in shape but not
similar enough to interchange without risk.
IjW^c\hZVihXdggZXianVii]ZWdiidb
IjW^c\YdZhcigZVX]i]ZWdiidb!^cigdYjX^c\YZVYkdajbZ
IjW^c\gZVX]Zhi]ZWdiidbWZ[dgZ[ZggjaZhZVih
Figure 71
Dead Volume created due to wrong Ferrule Type
KVaXd
G]ZdYncZ
EVg`Zg
HlV\Zad`
Figure 72
Ferrule Types from various Vendors
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Description and Installation
Troubleshooting
Avoiding Trapped Air Bubbles
It is common practice to make a connection only if the ports are wetted. The
liquid helps to prevent trapped air bubbles which are often very difficult to
remove or work out of a system.
Solvent Delivery Pumps are not priming
It has been observed that the solvent delivery pumps of the 'Pump Module' can
fail to prime at initial operation.
The pump has to be wetted at the solvent inlet port, or preferably prime the
tubing from the Solvent reservoir to the pump and make the connection.
Dry running the pump for a short time will not harm it.
Solvent Delivery Pump is not functioning
The pump flow rate depends greatly on the viscosity of the selected wash
solvent and the given backpressure of the entire system. The valve bore size
must be considered, as well as the tubing and injection loop internal diameter.
If in doubt about the delivered volume of a pump, a simple test can provide the
answer.
Execute the first test, only if the result is negative is it necessary to check
the flow from the pump directly.
Test 1: Pump flow delivered throughout the entire system
1 Prepare a graduated cylinder with a volume of 10 or 20 mL, Test solvent:
Water.
2 Select the following path from the local Terminal: Menu/Utilities/Wash
Station/select the desired Wash Station
The delivery pump and the DLW Actuator are activated, the delivered flow
can be collected at the DLW syringe needle.
Expected result: A volume of approximately 8 mL should collect within
30 s (Water, DLW syringe needle Gauge 22).
Test 2: Pump flow delivered at pump outlet
1 Disconnect the outlet tube from the pump module.
2 Connect PEEK tubing with the following dimensions to the outlet of the
pump module. Length 85 mm, ID 0.25 mm (0.010 inch).
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3 The wash solvent in the reservoir bottle should be water; if necessary,
replace the solvent.
4 Activate the pump by selecting following path:
Menu/F3-Setup/Objects/Events/Pwr-Out1 or Pwr-Out2.
5 Collect the pumped water in a graduated cylinder.
Within 30 s the pump should deliver approximately 10 mL.
This short test will give an indication of whether the system is performing as
expected. If none or very little flow is observed, perform a systematic check
using a process of elimination. Consult the table in “Troubleshooting” on
page 146 for more details.
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Troubleshooting
Verify the Functionality of the DLW Actuator/Solenoid
The DLW Actuator is electrically connected to the circuit of the heated syringe.
Increasing the heated syringe temperature activates the DLW Actuator. The
green LED is the indication that current is applied but please note that this is
not proof that the DLW Actuator opens the seat to allow a solvent flow. To
check the functionality, the following two tests are described:
Test 1: Electrical activation of the DLW Actuator
1 Select the DLW syringe using following path Menu | Utilities | Syringe | Standby
Temp | and select the standby temperature 65 °C.
Expected result: The green LED on the Syringe Holder assembly is turned
ON
In case this result is not observed, verify the electrical connection from the
DLW Actuator to the PCB.
NOTE
Do not reset the temperature if the second test is activated as well.
Test 2: Verify the seal tightness of the DLW Actuator
1 Close the DLW Actuator using the following path (from local Terminal):
Menu | Utilities | Syringe | Standby Temp | and select standby temperature
35 °C.
Expected result: The green LED on the Syringe Holder assembly is turned
OFF.
2 Place a beaker or other adequate vessel under the DLW Syringe Needle.
3 Select the Power-Out signal and activate both DLW Pumps. Path: Menu |
‘F3-Setup’ | Objects | Events | PwrOut1 for Pump1 , or ‘PwrOut2’ for Pump2
4 Activate both pumps by changing the status to the ‘Active State’ (Active
State OFF = Pump is activated, start. Active State ON = Pump is stopped).
Expected result: The DLW Actuator has been closed (Step 1). Activating the
DLW Pumps delivers solvent up to the point of the DLW Actuator. No
solvent should pass the DLW Actuator which verifies the tightness of the
seal.
NOTE
162
In case a solvent flow is observed during the test, remove the DLW Actuator and check if
particles are breaking the seal. If nothing special is observed, replace the device. The PNo.
is listed in the Appendix.
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Troubleshooting
Standard Chromatographic Tests
In this section standard chromatographic tests for 'carry-over' and
'repeatability' are described. These tests should provide basic information for
support technicians and users.
The tests described are examples only and should be adjusted accordingly if
the configuration of the HPLC system changes.
The test is designed for standard valves with a bore size of 0.25 mm, which
allows a flow rate range of 10 to 500 µL/min.
For both the carry-over and repeatability tests, conditions will be described
for UV detection.
Details are listed below. Note that the given levels to reach for carry-over and
repeatability do not reflect the specification values. The tests described here
should give the operator a confirmation whether the system is working as
expected. These tests are comparable to an OQ (Operational Qualification)
Test.
The test procedure is written for standard equipment, such as a standard
syringe of 100 µL and a loop size of 20 µL. The procedure does allow
configuring the system with different equipment, but the user has to bear in
mind certain limitations. For example, if a 100 µL syringe is installed
alongside a loop with a volume < 10 µL, the reliability of a partial loop
injection is questionable. The relative standard deviation will be above the
expected limit of 1.20 %. Example: 2 µL loop, 1 µL partial loop filling yields a
relative standard deviation of 2 – 4 %. Under such conditions, a full loop
injection is the only valid test.
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Carry-Over Test with UV-Detection
• Test Sample: Chlorhexidine, 0.6 mg/mL dissolved in Water : Acetonitrile =
90 : 10 + 0.1 % TFA
• Blank Solution: Water : Acetonitrile + 0.1 %TFA
• HPLC System:
• Binary Solvent Delivery Pump
• UV-Detector, Wavelength: 257 nm
• HPLC Column: Zorbax SB-C18, 2.1 x 50 mm; spherical 1.8 µm (or
equivalent)
• Column Temperature: Ambient
• Mobile Phase: A: Water + 0.1 % TFA; B: Acetonitrile + 0.1 % TFA
• Gradient:
• 10 % A hold for 0.5 min
• to 90 % B in 7.5 min
• hold for 1 min
• Wash Solvents:
• Wash1: Water + 0.1 % TFA
• Wash2: Acetonitrile + 0.1 % TFA
• Loop Size:
• 2 µL; Loop ID 0.13 mm
• Injection Valve: Agilent 1200 bar injection valve 5067-4123
• Cycle: DLW Standard
• Method Parameter Settings
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Table 33
Carry-Over Test with UV-Detection
Parameter
Value
Parameter
Value
Syringe
100 µL DLW
Pullup Del
500 ms
Sample Volume
7 µL
Inject to
Specified Inj.
Valve
Airgap Volume
3 µL
Inject Speed
5 µL/s
Front Volume
5 µL
Pre Inj Del
500 ms
Rear Volume
5 µL
Pst Inj Del
500 ms
Needle Gap Valve Clean
3 mm
Valve Clean Time Solvent 2
2s
Fill Speed
5 µL/s
Post Clean Time Solvent 2
2s
Fill Strokes
0
Valve Clean Time Solvent 1
2s
Stator Wash
0
Post Clean Time solvent 1
2s
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Repeatability Test with UV-Detection
• Test Sample
• Loop Overfill Caffeine, 520 mg/L
• Partial Loop Caffeine, 52 mg/L dissolved in Water : Acetonitrile = 90 :10
+ 0.1 % TFA
• HPLC System
• Binary Solvent Delivery Pump
• UV-Detector, Wavelength: 273 nm
• HPLC Column: Zorbax SB-C18, 2.1 x 50 mm; spherical 1.8 µm (or
equivalent)
• Column Temperature: Ambient
• Mobile Phase: Water : Acetonitrile = 90 :10 + 0.1 % TFA
• Gradient: isocratic for 2 min
• Wash Solvents:
• Wash1: Water + 0.1 % TFA
• Wash2: Acetonitrile + 0.1 % TFA
• Loop Size: 2 µL (Loop overfill, 7 µL); Loop ID: 0.13 mm; 20 µL (partial Loop,
10 µL); Loop ID 0.25 mm
• Injection Valve: Agilent 1200 bar injection valve 5067-4123
• Method Parameter Settings
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Table 34
Repeatability Test with UV-Detection
Parameter
Value
Parameter
Value
Syringe
100 µL DLW
Pullup Del
500 ms
Sample Volume
7 µL/10 µL
Inject to
Specified Inj.
Valve
Airgap Volume
3 µL
Inject Speed
5 µL/s
Front Volume
5 µL
Pre Inj Del
500 ms
Rear Volume
5 µL
Pst Inj Del
500 ms
Needle Gap Valve Clean
3 mm
Valve Clean Time Solvent 2
2s
Fill Speed
5 µL/s
Post Clean Time Solvent 2
2s
Fill Strokes
0
Valve Clean Time Solvent 1
2s
Stator Wash
0
Post Clean Time solvent 1
2s
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Description and Installation
Troubleshooting
Carry-Over and Repeatability Tests with UV-Detection
• Test Sample:
• Benzophenone 50 µg/mL dissolved in Water : Methanol = 50 : 50
• Blank Solution:
• Water : Methanol = 20 : 80
• HPLC System:
• Binary or isocratic Solvent Delivery Pump
• UV-Detector, Wavelength: 258 nm (10 mm path length)
• HPLC Column: C18 , 2.1 x50 – 100 mm spherical 2.7 – 5.0 µm (or
equivalent)
• Column Temperature: Ambient to 30 °C
• Mobile Phase: A: 20 % Water; B: 80 % Methanol
• Isocratic: 0.5 – 1.0 mL/min
• Wash Solvents:
• Wash1: Water : Methanol = 50 : 50
• Wash2: Water : Methanol = 50 : 50
• Loop Size:
• 20 µL; Loop ID 0.25 mm
• Injection Valve:
• Agilent 1200 bar injection valve 5067-4123
• Cycle:
• DLW Standard
• Method Parameter Settings
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Table 35
Test with Benzophenone and UV-detection
Parameter
Value
Parameter
Value
Syringe
100 µLDLW
Pullup Del
5000 ms
Sample Volume
60 µL
Inject to
Specified Inj.
Valve
Airgap Volume
3 µL
Inject Speed
5 µL/s
Front Volume
5 µL
Pre Inj Del
500 ms
Rear Volume
5 µL
Pst Inj Del
500 ms
Needle Gap Valve Clean
3 mm
Valve Clean Time Solvent 2
6s
Fill Speed
2 µL/s
Post Clean Time Solvent 2
6s
Fill Strokes
0
Valve Clean Time Solvent 1
6s
Stator Wash
0
Post Clean Time solvent 1
6s
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Description and Installation
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Replacing Parts
Warnings and Cautions
WA R N I N G
High voltage
Capacitors inside the instrument may still remain charged even if the instrument is
turned off.
➔ Always disconnect the power cord(s) from the power supply or from the various
power supplies if optional devices are installed before attempting any type of
maintenance.
➔ To avoid damaging electrical parts, do not disconnect an electrical assembly while
power is applied to the PAL system. Once the power is turned Off, wait
approximately 30 seconds before you disconnect an assembly.
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Replacing Parts
Replacing Control Board
Parts required
NOTE
#
p/n
Description
1
G6500-64220
PCB, Main for G427xA, G6501B, G6509B, AGI2
The Control board combines the MOTIO and the CPU boards in a single control board. This
board is required for the HTS System and operates only at Firmware level 4.0.X.
1 Follow the sequence shown in Figure 73 on page 172 to detach and release
the Control board from its position inside the X-axis.
a As shown, remove the dummy cover first.
2 Carefully pull the two ribbon cable connectors upwards to free them from
the board.
3 Install the replacement board in the reverse order.
NOTE
After reconnecting the two ribbon cables to the board, ensure that the cables are folded flat
and can slide inside the X-axis without damage.
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Replacing Parts
Figure 73
172
Replacing Control Board (HTS)
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Description and Installation
Replacing Parts
Injection Valve / Valve Rotor
1 After the mechanical connection of the PAL Valve Drive to the X-axis and
electrical connection to AUX1 at the board, turn on the power of the PAL
System.
The Firmware Object for the Valve type PAL VlvDrv-5067-4123_1 is preloaded,
the Valve Drive will go into the Standby position.
2 Insert the valve body into the shaft until the pin engages in the shaft.
3 Center the valve body by turning the valve body into position until the
vertical port is aligned.
4 Insert the Valve Body completely into the shaft until the pin on top of the
valve engages in the threaded Valve Drive Bracket. Tighten the Valve Body
with the polymer connecting nut.
HiVcYWndg^c_ZXi0"(%•
Figure 74
Valve Installation
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Description and Installation
Replacing Parts
Injection Unit
Follow the sequence of steps shown in “Assembling the LC Injector
HTC/HTS” on page 55.
DLW Pump Module
The following parts (keywords) from the DLW Pump Module can be exchanged
individually:
p/n
Description
G4277-60510
DLW Pump Module
(re-order for one solvent channel)
G6500-88035
Solvent Bottle Transfer Line including Glass Filter Solvent Inlet, 40 µm pore size
G4277-60513
Wash Station cable to System
G4277-60511
Pump control PCB
G4277-60512
DLW Tubing Kit
DLW Wash Station
The following parts (keywords) from the DLW Wash Station Module can be
exchanged individually:
174
p/n
Description
G4277-60514
DLW Wash Station incl. Waste Tube
G4277-60515
Needle Guide Length Tool
G4270-67205
Waste Tube
G4277-60517
Waste Tube (Valve to Wash station)
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Description and Installation
Replacing Parts
DLW Syringe
The following parts (keywords) from the DLW Syringe assembly can be
exchanged individually:
p/n
Description
G4277-80120
DLW Syringe 100 µL
G4277-80122
DLW Syringe Plunger (for syringe 100 µL)
G4277-60509
DLW Plunger Holder
DLW Syringe Holder Assembly
The following parts (keywords) from the DLW Syringe Holder assembly can be
exchanged individually:
p/n
Description
G4277-60601
DLW Syringe Holder Assembly (complete assembly)
G4277-60604
DLW Needle Holder Assembly
G4277-60602
DLW Holding Loop (installed in the DLW Syringe Assembly)
G4277-60503
DLW Manifold
(Entire unit assembled, consisting of DLW Solvent Selector, DLW Syringe Adapter
with Perfluor O-ring installed.)
G4277-60505
DLW Actuator/Solenoid
G4277-60603
DLW Actuator Control PCB (incl. cable)
G4277-60507
DLW Syringe Holder Fixation Screw
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Description and Installation
Replacing Parts
Replacing the DLW-2 Holding Loop
The DLW-2 Holding Loop is a combined part with injection needle and holding
loop as one piece. The holding loop is made out of stainless steel and the
inside is passivated with acid.
After a certain period of use or if mechanical damage occurs (for example bent
needle) the holding loop may need to be replaced.
Acid passivation is often used in HPLC technique to remove substances which
have accumulated over time. Typically a 20 % aqueous solution of nitric acid or
phosphoric acid is used.
It is highly recommended to periodically (for example once a year or more
often if the application requires it) clean critical parts which are always in
contact with the sample solution, such as the holding or the injection loop.
Please note that tubing, nuts, ferrules or any other part in the HPLC system
flow path made out of PEEK or another polymer may have resistance
problems against concentrated mineral acids.
1 Remove the PAL DLW-2 Syringe Holder Assembly from
3 Push up the DLW Needle Holder Assembly (1) and click it
the Z-axis.
out of the DLW Needle Adapter Block (2).
2 Remove the DLW Syringe and check at same time the
condition of the glass barrel and plunger; check tightness
of the plunger.
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4 Unscrew the Needle Holder Block.
5 Unscrew the retaining nut which tightens the holding
loop to the DLW Manifold (1). Pull out the DLW Holding
Loop (2).
6 Pull down the DLW-2 Holding Loop and remove it from the 7 Installing a new DLW-2 Holding Loop: Install a new loop in
DLW Syringe Holder. Release the retaining nut from
needle (1). Move the holding loop down to be able to
disengage the lower part of the holding loop (Needle)
from DLW Needle Holder Assembly (2).
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Replacing Parts
DLW Option Spare Parts
Spare Parts Order Information for DLW-2 Option
NOTE
This section of Spare Parts is intended for the DLW-2 Option only.
For Spare Parts for the previous version, DLW Option, see “Spare Part Order Information for
DLW Option” on page 183
For upgrade path from ‘DLW’ to ‘DLW-2’, see “Upgrade Path to DLW-2 Option” on
page 185.
DLW Pump
Module
178
p/n
Description
G4277-60510
DLW Pump Kit (single module); incl. electrical connections; 24 V
G6500-88035
Aspiration Tube Kit, consisting of: 1 pc. Tube PFA 1 pc. Glass Filer, 40 µm pore size
1 pc. Plug with feed through hole
G4277-60513
Cable for -xt Wash Station round connector, length 130 cm
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Description and Installation
Replacing Parts
DLW Wash
Station
p/n
Description
G4277-60511
DLW Pump Control Module PCB Kit
G4277-60512
DLW Wash Station Tubing Kit, Consisting of: Tubing set, wire supports and
connections
p/n
Description
G4277-60514
DLW Wash Station Block, complete assembly (incl. centering plate)
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Description and Installation
Replacing Parts
p/n
Description
G4277-50515
DLW Wash Station Insert Kit, consisting of: 2 pcs Inserts 1 pc. Waste Connector 5
pcs. Dummy Plugs
G4270-67205
Tube Waste for Wash Station, length 2 m
p/n
Description
G4277-80120
Syringe for DLW Option, 100 µL gastight; Ø7.7 mm; Scale Length 60 mm; Thread ¼
- 28 UNF. Removable needle not included.
G4277-80122
Replacement Plunger for Syringe SYRC DLW-R.; pkg of 10.
G4277-60509
DLW Plunger Holder
DLW Syringe
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Replacing Parts
DLW Syringe
Holder Assembly
p/n
Description
G4277-60601
DLW-2 Syringe Holder Assembly; complete Assembly with Holding Loop and
100 µL Syringe installed
G4277-60602
Kit DLW-2 Holding Loop with Needle, Adapter mounted, including Retaining Nut
G4277-60503
DLW Manifold Kit, complete, assembled
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182
Description and Installation
Replacing Parts
p/n
Description
G4277-60505
PAL DLW Actuator/Solenoid Kit; incl. screws, without cable
G4277-60603
DLW-2 Actuator Control PCB Kit; incl. cable and screw set
G4277-60507
Syringe Holder Fixation Screw, pkg of 1
G4277-60516
DLW Needle Length Guide Tool, pkg of 1
G4277-60604
DLW-2 Needle Holder
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Replacing Parts
Spare Part Order Information for DLW Option
NOTE
This section of Spare Parts is specific to the DLW Option.
The listing below shows only the spare parts which are specific to the ‘DLW Option’. The
other parts are either identical to the ‘DLW-2 Option Spare Parts’ or are backwards
compatible.
The upgrade path from ‘DLW’ to ‘DLW-2’ is described under “Upgrade Path to DLW-2
Option” on page 185.
DLW Syringe
DLW Syringe
Holder Assembly
p/n
Description
G4277-80125
Needle Kit for DLW Option, Gauge 22 PST 3, length 51 mm (3 pcs. per pack); incl.
Needle Retaining Nut.
p/n
Description
G4277-60502
Kit DLW Holding Loop with Needle, Adapter mounted
G4277-60504
DLW Flow Diverter Pkg of 1
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184
Description and Installation
Replacing Parts
p/n
Description
G4277-60506
DLW Actuator Control PCB Kit; incl. cable and screw set.
‘DLW’ version not available as single spare part anymore. The ‘DLW-2’ version is
not backwards compatible. Select ‘Upgrade Holder’, PNo.: DLW-2UpgrdKit. See
“Upgrade Path to DLW-2 Option” on page 185.
G4277-60508
DLW Needle Holder Kit
‘DLW’ version not available as single spare part anymore. The ‘DLW-2’ version is
not backwards compatible. Select ‘Upgrade Holder’, PNo.: DLW-2UpgrdKit. See
“Upgrade Path to DLW-2 Option” on page 185.
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Replacing Parts
Upgrade Path for DLW Option to DLW-2 Option
General Considerations
As described in “Development of the PAL DLW Option” on page 64, the DLW
technique has undergone several development steps. The current version,
DLW-2 Option, contains several improvements:
• Holding Loop
The DLW-2 Holding Loop is one single piece from the needle to the loop.
This eliminates several tube connections, the replacement needle and the
Flow diverter.
The loop is made of high quality stainless steel and the inner surface is
passivated with acid.
• DLW Syringe Holder Back-plate
The DLW-2 Syringe Holder back-plate has changed in size and form to allow
a smoother replacement of the holding loop. The Actuator is supported
from the back side for greater mechanical stability.
• DLW Pump Assembly
The DLW-2 Pump assembly is newly equipped with a housing to protect the
electrical parts from solvent splashes.
Upgrade Path to DLW-2 Option
The following parts are not backwards compatible with the older DLW Option:
p/n
Description
G4277-60603
DLW Actuator Control PCB (incl. cable)
G4277-60604
DLW Needle Holder Assembly
If one of these parts has to be replaced, is an upgrade kit available.
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Description and Installation
Replacing Parts
This kit is based on the DLW-2 version of syringe holder plate with installed
Actuator PCB and Needle Holder.
p/n
Description
G4277-60605
PAL DLW-2 Upgrade Kit Contains: DLW-2 Syringe Holder Plate, PAL DLW Actuator
Control PCB, PAL DLW Needle Holder
If one wishes to upgrade the entire DLW Holder to take advantage of the
mechanical changes and the stainless steel holding loop, is recommended to
use following order information:
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Description and Installation
Replacing Parts
p/n
Description
G4277-60601
DLW-2 Syringe Holder Assembly; complete Assembly with Holding Loop and
100 µL Syringe installed.
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Replacing Parts
Comparison DLW and DLW-2 Option
The following table compares the critical parts or parameters of the ‘DLW’ and
the ‘DLW-2’ Options. The specifications for the parts of the DLW-2 Option are
listed in “Specifications” on page 48.
Table 36
188
Comparison Parts and Parameters of the ‘DLW’ and ‘DLW-2’ Options
Part or Parameter
DLW Option
DLW-2 Option
Holding Loop
Material
OD
ID
Length
Volume
FEP Tubing
1/16” (1.58 mm)
0.50 mm
550 mm
108 µL
Stainless Steel, passivated
0.72 mm (Gauge 22)
0.41 mm
855 mm
118 µL
Syringe Needle
Length
Gauge
Material
51 mm
22
Stainless steel
Not applicable, integrated in
DLW-2 Holding Loop
DLW Flow Diverter
Length
OD
ID
Material
20.5 mm
0.72 mm (Gauge 22)
0.41 mm
Stainless Steel
Not applicable, integrated in
DLW-2 Holding Loop
DLW Internal Volume
Total Delay Volume
205 µL + Injection Loop volume
(204.7)
Manifold: 90 µL
Holding Loop : 108 µL
Syringe Needle: 6.7 µL
Installed Injection Loop
208 µL + Injection Loop volume
Manifold: 90 µL
Holding Loop: 118 µL
Installed Injection Loop
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Description and Installation
Maintaining the LC Injector HTC/HTS System
Maintaining the LC Injector HTC/HTS System
Regularly performing maintenance helps ensure accuracy and precision of the
autosampler System. Suggested intervals for maintenance procedures are
given below to ensure smooth, uninterrupted operation. If you use the
autosampler system extensively (for example, throughout nights and
weekends), or if you use corrosive solvents, you may need to perform the
maintenance procedure more frequently.
Table 37
Maintenance Checklist
Maintenance Step
Interval
Clean the outside of the instrument. Use only a
soft lint-free cloth dampened with mild soap and
water.
Weekly or as needed
Clean Instrument, syringe and surfaces
Weekly or as needed.
Replace the syringe plunger
The Syringe Plunger for a gas tight syringe
(polymer tip) has to be replaced on a regular
basis. The interval is highly dependent on the
application, throughput, quality of sample
solution (particles, etc.) and washing solvent.
It is advisable to check the tightness of the
plunger on a regular basis to gain experience
with the application.
Replace the Valve Needle Seal
Check the tightness of the needle in the needle
seal. See “Valve Needle Guide” on page 112.
Replace the Valve Rotor
Annually or more often, depending on the
throughput and quality of sample solution and
mobile phase.
Wash and Waste Solvent reservoirs.
Check flow restriction of frits.
Check quality of solvents, presence of biological
growth.
Weekly or as needed.
Replace the Septum as well to avoid carry-over.
Prime the entire DLW system by using the
‘Utilities’ functions ‘Wash Stations’ or
‘Syringe’.
Daily or as needed. Required before routine
running.
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Description and Installation
Maintaining the LC Injector HTC/HTS System
Table 37
Maintenance Checklist
Maintenance Step
Interval
Replace the DLW Syringe plunger.1
The Syringe Plunger for a gas tight syringe
(polymer tip) has to be replaced on a regular
basis. The interval is highly dependent on the
application, throughput, quality of sample
solution (particles, etc.) and washing solvent.
NOTE
It is advisable to check the tightness of the
plunger on a regular basis to gain
experience with the application.
Replace the DLW Syringe Needle (in one piece
with DLW-2 Holding Loop).
Only as needed (e.g. if bent).
Check all connections for tightness.1
Weekly or as needed.
Replace the DLW Holding Loop.1
Recommended to be changed if an excessive
carry-over effect is observed and cleaning of the
Holding Loop does not improve the situation.
Check the spring loaded Needle Holder
Assembly for functionality. Pay special attention
to the spring compression. 1
Weekly or as needed.
Check the functionality of the DLW
Actuator/Solenoid.1
Weekly or as needed.
Recommended to be changed once a year as a
preventative maintenance step. See “Verify the
Functionality of the DLW Actuator/Solenoid” on
page 162
1
NOTE
190
Dedicated to the DLW Option
There are no operator-serviceable or replaceable parts inside the power supply(ies) or the
autosampler System. In case of any failure, contact a representative of Agilent
Technologies.
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Appendices
Definition of Terms 192
Conventions of Naming 195
Injection Cycles 196
“LC-Inj” Cycle 196
DLW Principles and Operating Details 200
Priming the Solvent Lines 200
Location of Solvent and Waste Bottles 201
Functionality of the DLW Option 202
ICC Cycles used for the DLW Option 204
DLW Cycle Step-by-Step 211
Software Flow Chart 246
LC Injector HTC/HTS-xt System Flow Chart based on PAL Firmware
version 4.1.X (HTC) 246
PAL HTS-xt System Flow Chart based on PAL Firmware Level
4.1.X 250
External Connectors 253
Connector SER1 253
Connector TERMINAL or SER3 254
Connector SER2 255
Connector INTERFACE 1* 256
Connector INTERFACE 2 257
Connector AUX1* 258
Connector AUX2 259
Connector Wash Station 260
Connector Power 260
Connector MODBUS 261
Connector LAN 261
Connector Fuse 262
This chapter provides addition information.
Agilent Technologies
191
4
Appendices
Definition of Terms
Definition of Terms
Job Queue
A Job Queue is a list of sample processing Jobs. Jobs are executed in the order
displayed on the JOB QUEUE menu screens. New Jobs may be added to the
queue while samples are being processed.
Job
A Job contains the information needed by the LC Injector HTC/HTS to process
multiple samples by the same processing steps. The elements of a Job are a
Method and a Tray that define the location of the samples to be processed. For
identification, Jobs are automatically numbered from 01 to 99 and then
restarting with 01 when they are added to the Job Queue.
Cycle
A Cycle consists of the specific operations necessary to process one sample.
The Cycle operations are repeated for each sample within a Job. Cycles are
designed for specific applications.
Method
A Method defines how the samples are processed. The elements of a Method
are a Cycle, a Syringe and a Parameter List. Methods have names with up to
eight characters and can be edited, copied, and deleted.
Method Parameters
Method Parameters are associated with the Cycle operations. User-assigned
Parameter values define how a processing operation is performed. A zero
Parameter value will disable a Cycle operation. Cycle Parameters are
application-specific.
Tray Holder
A Tray Holder can hold one or more trays. Each Tray Holder has a reference
position (X-, Y-, Z-coordinates) that defines its location.
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Appendices
Definition of Terms
Tray
A Tray holds multiple samples. Trays are defined by designating the Tray Type
(see below) and the Tray Holder. Tray names are used to identify the sample
source within a Job.
Tray Type
A Tray Type defines the pattern and sampling sequence of sample locations
within a Tray.
Stack
A Stack is a particular type of Tray Holder that is designed to hold
micro-plates. A six-drawer Stack holds 12 standard microplates, two in each
drawer.
A three-drawer Stack holds six deep-well micro-plates, two in each drawer.
Object Manager
Software to load an Object List to an instrument if a Module (hardware
module) has been added to the System. In a special mode Object Manager can
also be used to create and maintain Object Lists.
Object List
If a Module (hardware) is added to an instrument, several Objects have to be
loaded into the firmware. These Objects are collected in an Object List and
stored in a file with the extension “*.pol”.
Object Lists are delivered together with Object Manager Software and are
grouped into folders for the different kind of Modules (e.g. Syringes, Tray
Holders, Valve Drives). The name of an Object List starts with the Module part
number with variants added (e.g. first or second Stack). The name of the root
folder includes the revision which is dependent on the firmware version (e.g.
“Object Lists Rev. K“ for firmware 2.X and 3.X).
Object Class
Each Object belongs to an Object Class (e.g. Syringes, Trays, Injectors).
The Object Class defines the Items of an Object.
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Appendices
Definition of Terms
Object Item
An Object contains several Items which can be numerical values with a
physical unit (e.g. X-, Y-, Z-Position, Penetration, Syringe Scale Length, Syringe
Volume) or references to other objects.
NOTE
The term “Parameter” is reserved for “ATOM Parameter” (Firmware commands to be used
for a Cycle).
Objects
Objects are data structures describing the properties of physical modules.
Certain modules (e.g. a Stack) require several objects.
Module
Hardware module, either part of a standard system configuration (e.g. Agilent
LC Injectors HTC/HTS, Agilent GC Sampler 80/120) or an optional addition
(e.g. SPME Upgrade for Agilent GC Injector, MT/DW Tray, Stack, and Cooler
Upgrades for Agilent LC Injectors HTC/HTS ). The term “Module” is
intentionally used to differentiate from “Object”, which is reserved for the
Firmware Object.
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Appendices
Conventions of Naming
4
Conventions of Naming
This section recommends standard naming convention for LC Injector
HTC/HTS Trays, Tray Types, and Tray holders. Following these conventions
will allow LC Injector HTC/HTS to be pre-configured for certain applications,
will simplify software backups and application development, and will improve
technical support and training.
Table 38
Naming Convention
Tray Type
Tray Description
VT200
Vial Tray, 200 positions (10 x 20)
For 7 mm micro-vials, 1 ml
VT98
Vial Trays, 98 positions (7 x 14)
For 12 mm vials, 2 ml
VT78
Vial Tray, 78 positions (6 x 13)
For 7 mm micro-vials, 1 ml
(opposite side of 98 positions Tray)
VT54
Vial Tray, 54 positions (6 x 9)
For 12 mm vials, 2 ml
VT21
Vial Tray, 21 Positions (7 x 14)
For 12 mm vials, 2 ml
(opposite side of 32 positions Tray)
VT32-10
Vial Tray, 32 positions (4 x 8)
For 23 mm headspace vials, 10 ml
VT32-20
Vial Tray, 32 positions (4 x 8)
For 23 mm headspace vials, 20 ml
MT96
Standard 96-position shallow microplate
DW96
Deep well 96-position microplate
MT384
High density 384-position shallow microplate
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Appendices
Injection Cycles
Injection Cycles
“LC-Inj” Cycle
No.
Processing Step
1
Reference X, Y, Z
axes and syringe
plunger
2
Wait for start signal
Start
(Sync Signals)
3
Read Barcode if
enabled
TRAY
(Tray Type)
Barcode ?
None
4
Syringe wash strokes
with solvent from
Wash1
Pre Cln Slv1
SYRINGE
(Syringes)
Wash1
(Wash
Stations)
Fill Speed
Eject Speed
Needle Penetr
Clean Volume
Max Fill Speed
Max Eject
Speed
Syringe wash strokes
with solvent from
Wash2
Pre Cln Slv2
SYRINGE
(Syringes)
Wash2
(Wash
Stations)
Rinse syringe with
sample selected
number of times
Pre Cln Spl
Sample
Volume
Fill Speed
Pullup Delay
SYRINGE
(Syringes)
TRAY
Waste
(Tray Type)
(Injectors)
5
6
196
Method
Parameter
Object
(Class)
Used Object
Value
(System)
Start Ref = ON
Fill Speed
Eject Speed
Needle Penetr
Clean Volume
Max Fill Speed
Max Eject
Speed
Eject Speed
Fill Volume
Needle Penetr
Needle Penetr
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Appendices
Injection Cycles
No.
Processing Step
Method
Parameter
Object
(Class)
Used Object
Value
7
Fill syringe with
sample using several
filling strokes;
aspirate air outside of
vial if Air Volume > 0
Sample
Volume
Air Volume
Fill Speed
Fill Strokes
Pullup Del
SYRINGE
(Syringes)
TRAY
(Tray Type)
Eject Speed
Overfill
Fill Volume
Needle Penetr
8
Move to injection
valve
Inject to
9
Wait for Inject signal
Inject
(Sync Signals)
10
Drive needle to
penetration depth
Inject to
11
Delay
Pre Inj Del
12
Switch valve to
Active position
13
Activate Injected
signal
14
Activate Injectd+
Signal
15
Dispense syringe
contents into sample
loop
Inject Speed
16
Delay
Pst Inj Del
17
Switch valve to
Standby position
18
Retract needle from
injector
19
Clean syringe and
needle with solvent
from Wash1
Pst Cln Slv1
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
(Injectors)
Needle Penetr
Injected
(Out Signals)
Pulse Time
Injectd+
(Out Signals)
Pulse Time
SYRINGE
(Syringes)
Wash1
(Wash
Stations)
Fill Speed
Eject Speed
Needle Penetr
Clean Volume
Max Fill Speed
Max Eject
Speed
197
4
Appendices
Injection Cycles
No.
Processing Step
Method
Parameter
Object
(Class)
Used Object
Value
20
Clean syringe and
needle with solvent
from Wash2
Pst Cln Slv2
SYRINGE
(Syringes)
Wash2
(Wash
Stations)
Fill Speed
Eject Speed
Needle Penetr
Clean Volume
Max Fill Speed
Max Eject
Speed
Rinse valve inlet with
solvent from Wash1
Vlv Cln Slv1
SYRINGE
(Syringes)
Wash1
(Wash
Stations)
SYRINGE
(Syringes)
Wash1
(Wash
Stations)
Home
(Positions)
21
22
23
198
Rinse valve inlet with
solvent from Wash2
Move to Home
position
Fill Speed
Eject Speed
Needle Penetr
Clean Volume
Max Fill Speed
Max Eject
Speed
Fill Speed
Eject Speed
Needle Penetr
Clean Volume
Max Fill Speed
Max Eject
Speed
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
4
Appendices
Injection Cycles
“LC-Inj” Cycle Parameter Description
Parameter
Description
CYCLE
LC-Inj Cycle
SYRINGE
Selected syringe
Sample Volume
Selected sample volume
Air Volume
Volume aspirated after the syringe needle is moved out of the sample liquid
Pre Cln Slv1
Pre injection syringe wash strokes with solvent from Wash1
Pre Cln Slv2
Pre injection syringe wash strokes with solvent from Wash2
Pre Cln Spl
Pre injection syringe rinse cycles with sample
Fill Speed
Speed of plunger movement used to aspirate sample
Fill Strokes
Number of filling strokes to aspirate sample
Pullup Del
Delay time between sample pull-up and ejection
Inject to
Name of Injector used for injection
Inject Speed
Speed of plunger movement used during sample injection
Pre Inj Del
Delay time prior sample injection
Pst Inj Del
Delay time after sample injection
Pst Cln Slv1
Post injection syringe wash strokes with solvent from Wash1
Pst Cln Slv2
Post injection syringe wash strokes with solvent from Wash2
Vlv Cln Slv1
Valve rinse cycles with solvent from Wash1
Vlv Cln Slv2
Valve rinse cycles with solvent from Wash2
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4
Appendices
DLW Principles and Operating Details
DLW Principles and Operating Details
Priming the Solvent Lines
The priming of the DLW Option is conveniently done directly from the local
terminal. There are several tasks which can be executed from the ‘Utilities’
section. It is recommended to use the function of the Wash station first,
followed by ‘Clean Syringe’.
Follow this description step-by-step to prime the solvent lines and the entire
DLW system. Solvent degassing and filtering is recommended. Important is to
observe the sequence of steps to ensure that Solvent 1 will be the last solvent
in the DLW System. Good practice is to have the wash solvent with lower
chromatographic strength (elution power) defined as ‘Wash Solvent 1’ and
‘Wash Solvent 2’ consistently as the solvent with a stronger elution power.
NOTE
For trouble-free DLW operation, make sure the two solvent lines are free of air bubbles at
all times. If the solvent lines are being connected for the first time or during a solvent
change, it is necessary to prime the solvent lines properly until air bubbles are no longer
visible. Solvent degassing and filtering is recommended.
Priming with Utilities Function ‘Wash Station’
Prime the PAL DLW System with the Utilities function ‘Wash Station’
1 Select Menu > Utilities > Wash Station.
2 Select Wash 2 > Function Key ‘F3’ > 'Move to Wash’ > Function Key ‘F2’ > 'Activate Valve’
3 Observe the green LED as an indicator that the DLW Actuator is activated.
Prime with Solvent 2 until no further air bubbles are observed.
4 Repeat the steps to prime with Solvent 1, by activating Wash 1.
Priming with Utilities Function ‘Syringe’
As second priming step for the PAL DLW System, the Utilities function
‘Syringe’ is recommended. This function allows control of the plunger
movement.
NOTE
200
It is important that the priming through ‘Wash Station’ is completed and that the solvent
lines are completely free of air bubbles.
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4
Appendices
DLW Principles and Operating Details
1 Select Menu > Utilities > Syringe > Function Key ‘F2’ ‘Cln Syr’.
2 Select Wash 2
a Select teh desired 'Clean Count' (recommended a minimum of 3 counts)
3 Observe the green LED as an indicator that the DLW Actuator is activated
and ensure that the plunger moves smoothly. Prime with Solvent 2 until no
further air bubbles are observed.
4 Repeat the steps to prime with Solvent 1, by activating Wash 1.
Other Priming functions or Troubleshooting
Another priming function is available in the section Utilities > Injector.
1 Select ‘Clean Injector’
2 Follow the dialog with similar logic as described under “Priming the Solvent
Lines” on page 200
In case any problems are observed, follow the instructions given in
“Troubleshooting” on page 146. Dedicated tests to verify the DLW
functionality are described in this section.
Location of Solvent and Waste Bottles
The DLW Option contains self-priming membrane pumps. The solvent bottles
can be placed either in the Fast Wash Station Holder or on the lab bench.
The Waste bottle must be placed >30 cm below the injection valve. Make sure
that the waste liquid can flow into the waste bottle without restriction. The
waste tubing must be placed above the level of the liquid. Ideally, the tube
should be fixed at the neck of the waste bottle.
NOTE
Use good lab practice to avoid contaminating wash solvents and bottles. Avoid the growth
of biological contaminants in pure water by either replacing it regularly or adding a small
amount of organic solvent, such as methanol or acetonitrile. Certain buffer solutions can
decompose at room temperature when exposed to light. Filtering the wash solvents before
filling the bottles, especially if salt buffers are used, is mandatory to avoid clogging of the
solvent paths.
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4
Appendices
DLW Principles and Operating Details
Functionality of the DLW Option
DLW Pumps
From the control point of view, the DLW pumps respond in the same manner
as the Fast Wash Station. The pumps are activated by 'Power-out Signals'. The
electric current setting for the DLW is different; this means that the
corresponding Firmware Objects have to be loaded for each DLW wash station
type. See “Loading the LC Injector HTC/HTS DLW Object” on page 98 for
further details.
The wetted parts in the pump are made of the following materials:
• Membrane: Kalrez (FFPM)
• Body, Valves: Ryton PPS
The pumps are self priming with a suction lift of up to 3 m water column.
DLW Actuator/Solenoid
The 'DLW Actuator/Solenoid' separates and completely shuts off the lines in the
direction of the syringe (sample loading) and the wash solvent.
After opening the DLW Actuator/Solenoid for the wash solvent lines, the
desired wash solvent can be pumped into the system by activating the
corresponding DLW pump.
The schematic in Figure 75 on page 203 illustrates this function.
The wetted parts in the DLW Actuator Solenoid are made of the following
materials:
• Solenoid body: PEEK
• Seal material: FFKM (Simriz)
202
NOTE
PEEK exhibits excellent chemical resistance to most commonly used chemicals. However,
the following solvents are not recommended for use with PEEK, DMSO, THF, methylene
chloride (dichloromethane), nitric acid, sulfuric acid. For more details see the compatibility
tables provided by the manufacturer of PEEK materials or components.
NOTE
In cases of defects, such as a defective or clogged Actuator/Solenoid, the LED would light
up but this does not mean that the solvent path is opened.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
Figure 75
DLW Manifold and Actuator/Solenoid
Figure 76
Schematic of DLW Manifold with Holding Loop
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4
Appendices
DLW Principles and Operating Details
ICC Cycles used for the DLW Option
The DLW Option can only be operated in combination with Agilent
ChemStation, Masshunter and EZChrom data systems.
For software control two cycles are provided. The first cycle is used for
Standard speed with optimized washing possibilities, and the second for a Fast
cycle with optimized throughput and less focus on carry-over (see Table 39 on
page 204).
The two cycles are explained in detail, see Table 40 on page 205 and Table 41
on page 208.
Table 39
DLW Cycle Definitions
Cycle Name
Cycle Description
LC-Inj DLW Standard
Standard injection cycle using all possibilities of the DLW Option.
The injection valve inlet port and the needle are washed with both
wash solvents (inside and out). An extra 'Stator Wash' can be
added for intensive washing of the injection valve (valve toggle).
LC-Inj Fast
The cycle is tuned for speed and high-throughput applications. The
difference with regard to the Standard DLW cycle is that some
steps are left out to shorten the cycle time. Details are explained
below.
General Considerations
The duration of the wash steps has to be established for each configuration
and application. The viscosity and surface tension of the individual wash
solvents composition and the backpressure of the system are factors to
consider.
Be aware that a higher backpressure builds up if the valve bore size or the
installed loop internal diameter are lower (standard valve bore 0.25 mm). The
standard loop internal diameter (ID) for loops provided with a volume of 5 ,
10 and 20 µl is 0.25 mm. The loop with 2 µl content volume has an ID of
0.125 mm.
Keep the tubing internal diameters of the tubing in line with the valve
dimensions, loop ID and flow rate.
204
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
Standard DLW Injection Cycle
Cycle Name: LC-Inj DLW Standard Rev03
Table 40
LC-Inj DLW Standard
Cycle Description
The System waits for the Sync Signal Ready
before the injection cycle is started.
Cycle Variable
NOTE
Sync Signal setting Start
Figure 77 on page 212
The Injection Valve is brought to a defined
position: Standby.
Inject to
Standby
The Rear air segment is pulled into the Holding
Loop.
Airgap Volume
Filling Speed
Figure 78 on page 213
The combined volumes of Rear-, Sample List-,
and Front-Volume are aspirated into the Holding
Loop.
Front Volume Figure 79 on page 214
Rear Volume
(SL.volume)
Airgap Volume
Filling Speed
Pullup Delay
Figure 80 on page 215
The Front air segment is aspirated.
The Injection Unit moves to the DLW Wash
station, position Wash1.
The needle is inserted (dipped) for 1 second to
wash the outer needle surface.
No plunger movement occurs at this step.
Figure 81 on page 216
The Injection Unit moves to the specified
Injection Valve.
The Front- and Airgap-Volume are ejected.
Inject to
Front Volume
Airgap Volume
Injection Speed
Figure 82 on page 217
The System waits for the signals from the data
system
Wait for DS
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4
Appendices
DLW Principles and Operating Details
Table 40
LC-Inj DLW Standard
Cycle Description
Cycle Variable
The Injection Valve is switched to position
Active.
The system pauses according to the delay time
Pre Inject Delay.
The loop is filled with the sample volume as
specified in the sample List.
The Injection Valve is switched to position
Standby, the loop contents are injected.
Timer 1 ( Delay Stator Wash is started and a
Start signal to the HPLC system is sent.
Inject to
Pre Injection Delay
Figure 83 on page 218
(SL.volume)
Injection Speed
Post Inject Delay
Timer 1
Figure 84 on page 219
The plunger of the DLW Syringe is pushed down
to dispense the Rear Sample and Air Segment
to Waste. The Holding Loop is still filled with
Wash Solvent 1.
(Syr. Eject Speed)
The DLW Actuator/Solenoid is activated to
deliver Wash Solvent 2 into the Holding Loop to
clean the Injection Valve from Port 1 to Port
Waste.
For this step the needle tip is lifted, releasing the
sealing pressure to enable rinsing around the tip
sealing point.
The DLW Actuator/Solenoid is activated,
the Wash Solvent (pump), the Needle Gap
and the Rinse Time are selectable.
Wash2
Inject to
Needle Gap Valve Clean
Valve Clean Time Solvent 2
Needle Gap is a parameter from Rinse Inj Atom.
the variable in this macro is: Needle Gap Valve
Clean. The function of this parameter is to lift
the needle in the injection port to allow rinsing
around the needle tip. This pressure of the
spring loaded balls in the DLW Syringe Holder
Assembly is released by moving approximately
3 mmup (default). This leaves a gap between the
needle tip and the valve bottom of approximately
1 mm, to enable a flush at this contact point.
Figure 86 on page 221
The Injection Unit is moved to the DLW Wash
Station, position Wash2.
The needle is rinsed inside and out with Wash
Solvent 2.
Wash2
(Syr.Eject Speed)
Post Clean Time Solvent 2
Figure 87 on page 222
The Injection Unit is moved back to the Injection
Valve. The Inlet Port and engraving to waste Port
is flushed with Wash Solvent 1 to prepare the
valve for the next injection.
Wash1
Inject to
Needle Gap Valve Clean
Valve Clean Time Solvent 1
Figure 88 on page 223
NOTE
The Atom Rinse Inj is now available
starting with Firmware 4.1.x.
206
Figure 85 on page 220
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
Table 40
4
LC-Inj DLW Standard
Cycle Description
Cycle Variable
The Injection Unit is moved back to the DLW
Wash Station, position Wash1 to flush the
syringe needle inside and out with Wash
Solvent 1.
This is a preparation step for the next injection.
Especially important for biofluid samples.
Wash1
Post Clean Time Solvent 1
Figure 89 on page 224
Cycle end for LC-Inj DLW Standard macro.
Figure 90 on page 225
An optional cleaning step is attached to the
DLW Standard injection cycle: Stator Wash or
Valve Toggle.
A Repeat-End loop can be activated with the
Count.
Stator Wash
Stator Wash count: 1
Stator Wash count: 1 = Cleaning step active
Stator Wash count: 0 = Cleaning step disabled
If Stator Wash is activated, the following steps
will be executed.
The Injection Unit is moved to the Injection
Valve. From the last step above, the Holding
Loop is filled with Wash Solvent 1.
Timer 1 is awaited to switch the valve (Toggle)
into Active position (fill loop).
Inject to
Delay Stator Wash
(Active)
“Stator Wash: Steps 1 – 2” on page 229
The DLW Actuator/Solenoid is activated to
deliver Wash Solvent 2 to the Holding Loop and
into the valve system.
The first solvent flush arriving at the valve is
Wash Solvent 1 followed by Wash Solvent 2.
Inject to
Wash2
Stator Wash Time Solvent 2
“Stator Wash: Step 3” on page 230
The Wash Solvent is changed to Wash Solvent
1.
Inject to
“Stator Wash: Step 4” on page 231
Wash1
Stator Wash Time Solvent 1
“Stator Wash: Step 5” on page 232
The Injection Valve is switched back to the
Standby position.
Inject to
(Standby)
“Stator Wash: Step 6” on page 233
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4
Appendices
DLW Principles and Operating Details
Fast DLW Injection Cycle
The “Fast” injection cycle differs from the “Standard” cycle in three points:
• The needle is not dipped in the Wash station “Wash1” after sample pickup
and before moving to the Injection Valve.
• The wash steps after injection are reduced to “Valve Clean” with Wash
Solvent 1 and Wash Solvent 2. The DLW needle is flushed in the DLW Wash
Station with “Wash Solvent 1” only.
• “Stator Wash” (Valve Toggle) is not available.
Cycle Name: LC-Inj Fast Rev03
Table 41
LC-Inj Fast Rev03
Cycle Description
Cycle Variable
The System waits first for the Sync Signal
Ready before the injection cycle is started.
NOTE
Sync Signal setting Start
Figure 98 on page 234
The Injection Valve is brought to a defined
position: Standby.
Inject to
Standby
The Rear air segment is pulled into the Holding
Loop.
Airgap Volume
Filling Speed
Figure 99 on page 235
The combined volumes of Rear-, Sample List-,
and Front-Volume are aspirated into the
Holding Loop.
Front Volume
Figure 100 on page 236
Rear Volume
(SL.volume)
Airgap Volume
Filling Speed
Pullup Delay
Figure 101 on page 237
The Front air segment is aspirated.
The Injection Unit moves to the specified
Injection Valve.
The Front- and Airgap-Volume is ejected to
Waste.
208
Inject to
Front Volume
Airgap Volume
Injection Speed
Figure 102 on page 238
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
4
Appendices
DLW Principles and Operating Details
Table 41
LC-Inj Fast Rev03
Cycle Description
Cycle Variable
The System waits for a signal from the data
system
Wait for DS
The Injection Valve is switched to position
Active.
The system pauses according to the delay time
Pre Inject Delay.
The loop is filled with the sample volume as
specified in the sample List.
The Injection Valve is switched to position
Standby, the loop contents are injected.
Inject to
Pre Injection Delay
(SL.volume)
Figure 103 on page 239
Injection Speed
Post Inject Delay
Figure 104 on page 240
The plunger of the DLW Syringe is pushed down
to dispense the Rear Sample and Air Segment
to Waste. The Holding Loop is still filled with
Wash Solvent 1,
Injection Speed
Figure 105 on page 241
The DLW Actuator/Solenoid is activated to
deliver Wash Solvent 2 into the Holding Loop to
clean the Injection Valve from Port 1 to Port
'Waste'.
For this step the needle tip is lifted, releasing the
sealing pressure to enable rinsing around the tip
sealing point.
Inject to
Wash2
Needle Gap Valve Clean
Valve Clean Time Solvent 2
Needle Gap is a parameter from ‘Rinse Inj’
Atom. The variable in this macro ’Needle Gap
Valve Clean’. The function of this parameter is
to lift the needle in the injection port to allow
rinsing around the needle tip. The pressure of
the spring-loaded balls in the DLW Syringe
Holder Assembly is released by moving approx 3
mm up (default). This leaves a gap between the
needle tip and the valve bottom of approx. 1 mm,
to enable a flush at this contact point.
NOTE
The Atom ‘Rinse Inj’ is new, available
starting with FW 4.1.X. The DLW
Actuator/Solenoid is activated, the Wash
Solvent (pump), the ‘Needle Gap’ and the
‘Rinse Time’ are selectable.
NOTE
The Needle Gap Valve Clean is explained
in detail in the macro DLW Priming.
Figure 106 on page 242
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4
Appendices
DLW Principles and Operating Details
Table 41
210
LC-Inj Fast Rev03
Cycle Description
Cycle Variable
Wash Solvent 1 follows to prepare the valve for
the next injection.
Wash1
Inject to
Needle Gap Valve Clean
Valve Clean Time Solvent 1
Figure 107 on page 243
The Injection Unit is moved back to the DLW
Wash station, position Wash1 to flush the
syringe needle inside and out with Wash
Solvent 1.
This is a preparation step for the next injection.
Especially important for biofluid samples.
Wash1
Post Clean Time Solvent 1
Figure 108 on page 244
Cycle end for LC-Inj DLW Fast macro.
Figure 109 on page 245
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
DLW Cycle Step-by-Step
NOTE
The following illustrations display a valve configuration with a loop mounted between port
3 and port 6. For the Agilent 1200 bar injection valve the general principles for the DLW
operation is the same, although the loop is mounted between port 2 and port 5.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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4
Appendices
DLW Principles and Operating Details
Standard Cycle
Standard: Cycle Start
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212
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Standard Injection Cycle Start
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
Standard: Step 1
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Figure 78
Standard Injection Cycle Step 1: Aspirate Rear Air Segment
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
213
4
Appendices
DLW Principles and Operating Details
Standard: Step 2
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214
Standard Injection Cycle Step 2: Get Sample Aspirate Rear, Inject and Front
Volume
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
4
Appendices
DLW Principles and Operating Details
Standard: Step 3 — 4
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Figure 80
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Standard Injection Cycle Step 3-4: Aspirate Front Air Segment
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
215
4
Appendices
DLW Principles and Operating Details
Standard: Step 5 — 6
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216
Ejbe
Standard Injection Cycle Step 5-6: Passive needle clean outside (dip) in wash
position 1
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
Standard: Step 7 — 8
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Figure 82
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Standard Injection Cycle Step 7-8: Dispense Front Air Segment and Front
Sample Volume to Waste
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
217
4
Appendices
DLW Principles and Operating Details
Standard: Step 9 — 10
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218
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Standard Injection Cycle Step 9-10: Valve is switched to LOAD position, Loop
is filled with 'Inject Volume'
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
4
Appendices
DLW Principles and Operating Details
Standard: Step 11
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Ejbe
Standard Injection Cycle Step 11: Valve is switched to INJECT position, start
chromatographic process
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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4
Appendices
DLW Principles and Operating Details
Standard: Step 12
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Figure 85
220
Ejbe
Standard Injection Cycle Step 12: Rear Sample Volume and Air Segment are
dispensed to Waste
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
Standard: Step 13 — 14
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Figure 86
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Standard Injection Cycle Step 13-14: Valve Clean with Wash Solvent 2
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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4
Appendices
DLW Principles and Operating Details
Standard: Step 15 — 16
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Figure 87
222
Ejbe
Standard Injection Cycle Step 15-16: Active Syringe Needle wash with Wash
Solvent 2
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
Standard: Step 17 — 18
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Standard Injection Cycle Step 17-18: Valve Clean with Wash Solvent 1
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
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4
Appendices
DLW Principles and Operating Details
Standard: Step 19 — 20
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224
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Standard Injection Cycle Step 19-20: Active Syringe Needle Wash with Wash
Solvent 1
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
Standard: Cycle End
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Standard Injection Cycle End
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
225
4
Appendices
DLW Principles and Operating Details
Additional Valve Toggle Step to the DLW Standard Cycle
Considerations for additional 'Stator Wash' Cleaning Step
The DLW Standard Cycle has the built-in option for the user to toggle the
injection valve at the end of the chromatographic run before equilibration of
the column to the start conditions. If the method variable ' Stator Wash' is set
to ' 1', the extra cleaning process for the valve, with ' Valve Toggle', becomes
part of the standard cycle.
If the Method variable is deactivated (setting ' 0'), the DLW Standard cycle
ends as shown in Figure 90 on page 225
The macro (cycle) is written such that the optional valve toggle steps can be
executed before re-equilibration of the column. The exact time the valve needs
to be switched on hast to be synchronized with the chromatographic method
by the user, Method variable ' Delay Stator Wash'. The two wash solvents are
timed by the Method variables ' Stator Wash Time Solvent 1' and ' Stator Wash
Time Solvent 2'. After these wash times have elapsed, the valve is switched back
to the start position.
6WhdgWVcXZ
7
Figure 91 on page 226illustrates the recommended retention time for ' Stator
Wash' or ' Valve Toggle' times.
KVakZid\\aZ
<gVY^Zci
HiVgi
Figure 91
I^bZ
Timing for “Stator Wash” Step
From the chromatographic point of view, it is important to understand the
optional cleaning step in detail. It is assumed that the valve stator between
226
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
4
Appendices
DLW Principles and Operating Details
ports 1 and 6 (example standard Cheminert Valve) is contaminated and
cannot be cleaned during the standard injection process. The valve toggle
brings the engraving back between the two ports. Flushing the valve with both
wash solvents eliminates any remaining sample material located between
stator ports 1 and 6.
What points need to be considered when using the ' Stator Wash'or ' Valve
Toggle' option?
• Observe the rules if biofluid samples are injected. The first sample contact
should always be with an aqueous solution to avoid protein precipitation.
After washing with an organic solvent (higher elution power) the system
must be flushed again with Wash Solvent 1.
• The first toggle near the end of the chromatographic cycle provides the
advantage that the sample loop is already flushed out, first using the mobile
phase with a solvent of high elution power (assuming gradient application).
• The second valve toggle time follows immediately after the second solvent
flush is completed. A second switching time cannot be programmed. The
waiting period before the second valve toggle should be long enough so that
the entire system is flushed out by both wash solvents.
Consider the entire delay volume to determine the second valve toggle time.
The DLW internal volumes are:
• Manifold, 90 µl,
• Holding Loop, 118 µl(inclusive Needle DLW-2 version,
• Installed Injection Loop.
Total delay volume: 208 µl+ Loop content volume.
• The second valve toggle (back to starting conditions) should be done before
the system equilibration time has started. The Loop contents should ideally
be a solvent of a low elution power when switched back.
• If isocratic chromatography is applied, it is possible that the remaining
contaminants are washed into the system and can build up higher
background noise for the column and/or detector over a longer period of
time.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
227
4
Appendices
DLW Principles and Operating Details
Additional Cleaning step 'Stator Wash' or 'Valve Toggle' Step-by-Step
Stator Wash: End of Standard Injection Cycle, Start for additional
Cleaning Step 'Valve Toggle'
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6XijVidgkVakZ
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Ejbe'
=daY^c\adde
8dajbc
HVbeaZ
Ejbe
Figure 92
228
Additional Cleaning Step: End of Standard Injection Cycle, Start for additional
Cleaning Step 'Valve Toggle'
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
Stator Wash: Steps 1 – 2
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
8dajbc
HVbeaZ
Ejbe
Figure 93
Additional Cleaning Step: Valve Switched to Load Position (toggle), Valve
clean with Wash Solvent 1
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
229
4
Appendices
DLW Principles and Operating Details
Stator Wash: Step 3
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
8dajbc
HVbeaZ
Ejbe
Figure 94
230
Additional Cleaning Step: Valve Clean with Wash Solvent 2
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
Stator Wash: Step 4
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
8dajbc
HVbeaZ
Ejbe
Figure 95
Additional Cleaning Step: Wash Solvent 2 dispensed by Wash Solvent 1
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
231
4
Appendices
DLW Principles and Operating Details
Stator Wash: Step 5
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
8dajbc
HVbeaZ
Ejbe
Figure 96
232
Additional Cleaning Step: Second Valve Clean with Wash Solvent 1
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
Stator Wash: Step 6
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
8dajbc
HVbeaZ
Ejbe
Figure 97
Additional Cleaning Step: Valve switched back to Inject Position (toggle)
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
233
4
Appendices
DLW Principles and Operating Details
Fast Injection Cycle Step-by-Step
Fast: Cycle Start
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
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=daY^c\adde
LVh]'
LVh]&
8dajbc
HVbeaZ
Ejbe
LVhiZ
Figure 98
234
Fast Injection Cycle: Start
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
Fast: Step 1
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
GZVgV^ghZ\bZci
LVh]'
LVh]&
8dajbc
HVbeaZ
LVhiZ
Figure 99
Ejbe
Fast Injection Cycle: Aspirate Rear Air Segment
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
235
4
Appendices
DLW Principles and Operating Details
Fast: Step 2
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
LVh]'
LVh]&
8dajbc
HVbeaZ
LVhiZ
Ejbe
Figure 100 Fast Injection Cycle: Get Sample, Aspirate Rear, Inject and Front Volume
236
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
Fast: Steps 3-4
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
;gdciV^ghZ\bZci
LVh]'
LVh]&
8dajbc
HVbeaZ
LVhiZ
Ejbe
Figure 101 Fast Injection Cycle: Aspirate Front Air Segment
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
237
4
Appendices
DLW Principles and Operating Details
Fast: Steps 5 - 6
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
;gdcikdajbZ
LVh]'
LVh]&
8dajbc
HVbeaZ
LVhiZ
Ejbe
Figure 102 Fast Injection Cycle: Dispense Font Air Segment and Front Sample Volume to
Waste
238
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
4
Appendices
DLW Principles and Operating Details
Fast: Steps 7 - 8
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
;gdcikdajbZ
>c_ZXikdajbZ
LVh]'
LVh]&
8dajbc
HVbeaZ
LVhiZ
Ejbe
Figure 103 Fast Injection Cycle: Valve is switched to LOAD position, Loop is filled with
'Inject Volume'
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
239
4
Appendices
DLW Principles and Operating Details
Fast: Step 9
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
LVh]'
LVh]&
8dajbc
HVbeaZ
LVhiZ
Ejbe
Figure 104 Fast Injection Cycle: Valve is switched to INJECT position, start
chromatographic process
240
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
4
Appendices
DLW Principles and Operating Details
Fast: Step 10
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
LVh]'
LVh]&
8dajbc
HVbeaZ
LVhiZ
Ejbe
Figure 105 Fast Injection Cycle: Rear Sample Volume and Air Segment are dispensed to
Waste
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
241
4
Appendices
DLW Principles and Operating Details
Fast: Steps 11 - 12
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
LVh]'
LVh]&
8dajbc
HVbeaZ
LVhiZ
Ejbe
Figure 106 Fast Injection Cycle: Valve Clean with Wash Solvent 2
242
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
Fast: Steps 13 - 14
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
LVh]'
LVh]&
8dajbc
HVbeaZ
LVhiZ
Ejbe
Figure 107 Fast Injection Cycle: Valve Clean with Wash Solvent 1
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
243
4
Appendices
DLW Principles and Operating Details
Fast: Step 15
Ejbe&
HdaZcd^Y$
6XijVidgkVakZ
Ejbe'
=daY^c\adde
LVh]'
LVh]&
8dajbc
HVbeaZ
LVhiZ
Ejbe
Figure 108 Fast Injection Cycle: Active Syringe Needle Wash with Wash Solvent 1
244
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
DLW Principles and Operating Details
4
Fast: Cycle End
HdaZcd^Y$
6XijVidgkVakZ
Ejbe&
Ejbe'
=daY^c\adde
LVh]'
LVh]&
8dajbc
HVbeaZ
LVhiZ
Ejbe
Figure 109 Fast Injection Cycle: End
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
245
4
Appendices
Software Flow Chart
Software Flow Chart
LC Injector HTC/HTS-xt System Flow Chart based on PAL
Firmware version 4.1.X (HTC)
NOTE
Firmware version 4.1.X., as shown in the flow chart, corresponds to the LC Injector
HTC/HTS-xt System. The changes from Firmware version 2.5.X., as used for the LC Injector
HTC/HTS System, are emphasized in this LC Injector HTC/HTS User Manual.
Figure 110 LC Injector HTC-xt Firmware Overview, Page 1
246
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
Software Flow Chart
4
Figure 111 LC Injector HTC-xt Firmware Overview, Page 2
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
247
4
Appendices
Software Flow Chart
Figure 112 LC Injector HTC-xt Firmware Overview, Page 3
248
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
Software Flow Chart
4
Figure 113 LC Injector HTC-xt Firmware Overview, Page 4
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
249
PAL HTS-xt System Flow Chart based on PAL Firmware Level 4.1.X
PAL HTS-xt System Flow Chart based on PAL Firmware Level 4.1.X
NOTE
Firmware version 4.1.X., as shown in the flow chart, corresponds to the PAL-xt System. The
changes from Firmware version 2.5.X., as used for the PAL System, are emphasized in this
PAL User Manual.
Figure 114 PAL HTS-xt Firmware Overview, Page 1
Figure 115 PAL HTS-xt Firmware Overview, Page 2
250
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
PAL HTS-xt System Flow Chart based on PAL Firmware Level 4.1.X
Figure 116 PAL HTS-xt Firmware Overview, page 3
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
251
PAL HTS-xt System Flow Chart based on PAL Firmware Level 4.1.X
Figure 117 PAL HTS-xt Firmware Overview, Page 4
252
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
External Connectors
4
External Connectors
Connector SER1
Connector SER1
1
Bridged with Pin7; Special grounding, do not alter.
2
Bridged with Pin 4; Do not alter.
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Pin
Signal Name
1
NC
2
RXD
3
TXD
4
DTR1
5
GND
6
NC
7
RTS2
8
NC
9
NC
253
4
Appendices
External Connectors
Connector TERMINAL or SER3
Connector TERMINAL or SER3
254
Pin
Signal Name
1
GND
2
RXD
3
TXD
4
NC
5
NC
6
+5 V
7
NC
8
GND
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
External Connectors
4
Connector SER2
Connector SER2
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Pin
Signal Name
1
GND
2
RXD
3
TXD
4
NC
5
NC
6
+5 V
7
NC
8
GND
255
4
Appendices
External Connectors
Connector INTERFACE 1*
*
APR Control-LC Injector HTC/HTS Board:”INTERFACE” and not
“INTERFACE1”
Connector INTERFACE 1
256
Pin
Signal Name
1
Pwr-Out1 +
2
Pwr-Out2 +
3
SW-Out1 N0
4
SW-Out1 COM
5
SW-Out2 N0
6
Opto-In1 +
7
TTL-In1 +
8
GND
9
Pwr-Out1 -
10
Pwr-Out2 -
11
TTL-In2
12
SW-Out2 COM
13
TTL-In3
14
Opto-In1 -
15
+5 V
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
External Connectors
4
Connector INTERFACE 2
Connector INTERFACE 2
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Pin
Signal Name
1
TTL-In1
2
TTL-In2
3
TTL-In3
4
TTL-Out1
5
TTL-Out2
6
TTL-Out3
7
Opto-In1 +
8
Opto-In2 +
9
SW-Out1 NO
10
SW-Out2 NO
11
Pwr-Out1 +
12
Pwr-Out2 +
13
+5V
14-19
GND
20
Opto-In1 –
21
Opto-In2 –
22
SW-Out1 COM
23
SW-Out2 COM
24
Pwr-Out1 –
25
Pwr-Out2 -
257
4
Appendices
External Connectors
Connector AUX1*
*
APRControl-LC Injector HTC/HTS Board: ”AUX” and not “AUX1”
Connector AUX1
258
Pin
Signal Name
1,2
Motor A1
11, 12
Motor A2
3, 4
Motor B1
13, 14
Motor B2
15, 18, 19
GND
6
Temp +
16
Temp -
7
Sens
17
+5 V
8, 9
Heater
10,20
+36 V
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Appendices
External Connectors
4
Connector AUX2
Connector AUX2
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Pin
Signal Name
1,2
Motor A1
11, 12
Motor A2
3, 4
Motor B1
13, 14
Motor B2
15, 18, 19
GND
6
Temp +
16
Temp -
7
Sens
17
+5 V
8, 9
Heater
10,20
+36 V
259
4
Appendices
External Connectors
Connector Wash Station
Connector WASHSTATION
Pin
Signal Name
1
Pwr-Out1+
2
Pwr-Out1-
3
Pwr-Out2+
4
Pwr-Out2-
Pin
Signal Name
1
+36 V
2
GND
3
NC
'ELG"Dji&" (ELG"Dji'
&ELG"Dji&
)ELG"Dji'"
Connector Power
Connector POWER
& (+K
(C8
'<C9
260
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
4
Appendices
External Connectors
Connector MODBUS
Connector MODBUS
Pin
Signal Name
1
TXD
2—7
+36 V
8
RXD
9 — 14
GND
Connector LAN
Connector LAN
A:9aZ[i
Pin
Signal Name
DIR
Primary Function
1
TX+
Out
Transmit Data +
2
TX-
Out
Transmit Data -
3
RX+
In
Receive Data +
4, 5
Not used
6
RX-
7, 8
Not used
A:9g^\]i
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Terminated
In
Receive Data Terminated
261
4
Appendices
External Connectors
Connector Fuse
262
Fuse Type
FST 5x20
Rating
6.3 A
Rated
250 VAC
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Index
Index
A
actuator/solenoid
DLW 202
air bubbles 160
AUX1
connector 258
AUX2
connector 259
B
bottles
solven 201
waste 201
C
carry-over test
uv-detection 164
carryover 150
chromatographic test 163
COM Port
settings 120
communication
computer 120
LAN 122
components
unpacking 54
connector
AUX1 258
AUX2 259
interface 1 256
interface 2 257
LAN 255
power 260
SER1 253
SER3 254
terminal 254
WASHSTATION 260
control terminal 17
cycle parameter
description 199
cycle
definitions 204
injection 196
D
delivery 66
detector signal
no 146
very low 146
dilutor
function key 35
item 35
DLW wash station
installing 71
E
electrical connections
LC Injectors HTC 86
single injection valve 83
electrical specifications 50
enter button 19
error message 146
extended user mode
access 126
F
f3-setup 127, 134
ferrule 159
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
firmware
4.1.X 246
flow chart 246
free items 41
free objects 41
function key
wash station 33
function key
dilutor 35
injector 32
syringe 28
tool 36
tray 30
vial 34
function keys 19
function
info 38
H
hardware
item 39
holding loop
replacing 176
I
inclination
correction 95
info function 38
injection cycle
fast 234
injection unit
collision 151
installing 56
injection valve
263
Index
installing 61
reference point 102
injection
cycle 196
injector
function key 32
item 32
installing
injection unit 56
keypad terminal 60
microplate stack 84
power supply 60
trayholder 85
interface 1
connector 256
interface 2 257
item
dilutor 35
hardware 39
injector 32
maintenance 40
software 39
tool 36
vial 34
wash station 33
items
syringes 29
tray 31
ESCape and STOP
K
keypad terminal
installing 60
264
19
L
LAN
communication 122
connector 255
loader program 118
logfile 37
M
maintenance 189
item 40
menu screens 18
methods
creating 20
delete 22
edit/view 22
microplate stack
installing 84
mobile phase
wrong ports 147
N
naming
convention 195
needle seal 112
nut 159
J
job queue
building and starting
job queue
aborting 26
restarting 26
job 23
P
keys
O
24
object
collision 152
DLW 98
manager 109
not detected 152
objects 44, 134
output signals 115
parts
spare 111
penetration value
definition 103
performance specification 49
physical specifications 51
pilot distance 156
plumbing 106
positions
actuator 110
valve 110
power supply
installing 60
pump holder
mounting 68
pump module
DLW 174
pumps
DLW 202
R
reference point
injection valve 102
waste 101
reference points
wash1/wash2 100
reference positions
defining 88
requirements
hardware 65
operating and environmental
software 65
re-use
nut/ferrule 155
52
S
safety information
sample flow
8
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
Index
wrong ports 147
sample
backing up 147
valve needle guide 147
scroll knob 19
service 132
settings
COM Port 120
setup functions 42
software
item 39
requirements 53
solvent delivery pump 160
solvent lines
priming 200
sound pressure level 52
sounds 43
special function 126
specifications
6-port Agilent injection valve
5067-4123 107
synchronization 115
syringe holder assembly
DLW 175
inserting 79
syringe
DLW 175
function key 28
items 29
system 129
overview 62, 47
T
terms
definition 192
test
repeatability 166
tightening
stepwise 154
time 43
tools 36
tool
function key 36
item 36
tray column
definition 94
tray holder 91
tray row
definition 94
tray type 138
trayholder
installing 85
trays 93, 142
tray
function key 30
items 31
troubleshooting 146
tube
cut 157
tubing
connection 153
item 33
WASHSTATION
connector 260
waste
reference point 101
U
utility functions
27
V
valve needle guide 112
valve toggle 226
valve drive 105
valve 105
vial
function key 34
item 34
W
wash station
DLW 174
fuction key 33
Agilent 1290 Infinity LC Injectors HTC/HTS User Manual
265
www.agilent.com
In This Book
The manual describes the following:
• Introduction
• Installing
• Operating
• Troubleshooting
• Maintenance
• Replacing Parts
• Limitations
• Appendices
Agilent Technologies 2009, 2011-2012
Printed in Germany
05/2012
*G4277-90000*
*G4277-90000*
G4277-90000 Rev. B
Agilent Technologies